WO2022190261A1 - Cell culture vessel - Google Patents

Cell culture vessel Download PDF

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Publication number
WO2022190261A1
WO2022190261A1 PCT/JP2021/009546 JP2021009546W WO2022190261A1 WO 2022190261 A1 WO2022190261 A1 WO 2022190261A1 JP 2021009546 W JP2021009546 W JP 2021009546W WO 2022190261 A1 WO2022190261 A1 WO 2022190261A1
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WO
WIPO (PCT)
Prior art keywords
cell culture
culture vessel
container body
tubular portion
bottom portion
Prior art date
Application number
PCT/JP2021/009546
Other languages
French (fr)
Japanese (ja)
Inventor
学司 加藤
順一 桑原
Original Assignee
株式会社サンプラテック
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 株式会社サンプラテック filed Critical 株式会社サンプラテック
Priority to PCT/JP2021/009546 priority Critical patent/WO2022190261A1/en
Priority to JP2023504964A priority patent/JPWO2022190261A1/ja
Publication of WO2022190261A1 publication Critical patent/WO2022190261A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • the present disclosure relates to a cell culture vessel for culturing cells, and in particular, to a cell culture vessel used for perfusion culture that automatically supplies culture medium at a constant low speed and discharges the same amount of culture medium at the same time.
  • culture vessels such as dishes (petri dishes), well plates, and flasks are widely used. Even if the dishes and well plates had lids, there was a gap just by putting them on, and it was impossible to seal them. Flasks can be hermetically sealed, but all of these conventional culture vessels require the opening and closing of lids for operations such as exchanging the culture solution. Therefore, these culture vessels are highly versatile for use in cell culture for research purposes. There was a problem that it was difficult to equalize the quality of cultured cells.
  • a culture vessel used in an automatic culture apparatus is provided with, for example, a pair of ports. These ports are provided, for example, in the lid of the culture vessel and extend downward from the lid to be arranged inside the culture vessel.
  • a new culture solution is supplied to the culture container through one port, and the culture solution inside the culture container is discharged to the outside through the other port.
  • the amount of culture medium supplied from one port and the amount of culture medium discharged from the other port are not exactly the same, the liquid in the container may dry up or overflow during long-term cultivation. Become.
  • one pump is used for one container, and the culture solution is supplied from one port under positive pressure and discharged from the other port by overflowing, or A method in which the culture medium is discharged from the port under negative pressure and the culture medium is drawn in from the other port is effective. If this is done all at once, turbulence will occur and the cells will be damaged. Therefore, perfusion culture is performed in which the culture solution is automatically supplied at a constant low speed and the same amount of culture solution is discharged at the same time. In this perfusion culture, in order to realize smooth liquid flow, it is important to have a closed system and to fill the entire interior of the culture vessel with the culture medium without creating a gas layer or air bubbles.
  • the present disclosure has been conceived under such circumstances, and a main object of the present disclosure is to provide a cell culture vessel suitable for improving the inconvenience of performing perfusion culture in a closed system. .
  • a cell culture vessel includes a vessel body having an opening at one end in a first direction and having a first cylindrical portion extending in the first direction, and a plug member capable of closing the opening. and, the plug member includes a second cylindrical portion that abuts and seals the inner peripheral surface of the first cylindrical portion, and a diameter from the one side end of the second cylindrical portion in the first direction. a first flange portion that extends outward in the direction and closes the one side end of the first tubular portion in the first direction; a plate-like first bottom part that extends in the direction of the first direction and closes the inside of the second cylindrical part when viewed in the first direction, and the first bottom part has the one side end in the first direction that extends to the outside.
  • a plurality of ports each having a flow path that opens to the first direction and that the other side end in the first direction opens in the inner space of the container body, and the first bottom portion is provided in the first direction a ceiling surface facing the other side, a ceiling recess recessed from the ceiling surface toward the one side in the first direction is formed in the first bottom portion, and the ceiling recess is one of the plurality of ports; Communicate with at least one of the channels.
  • the ceiling recess communicates with the outer peripheral edge of the first bottom.
  • annular top plate portion a third tubular portion extending from the outer peripheral edge of the top plate portion in the thickness direction of the top plate portion and fitted onto the first tubular portion, At least one of the container body and the outer lid has the first tubular portion and the third flange portion in contact with the first tubular portion. Locking means are provided to prevent relative movement with the tubular portion.
  • the plug member is made of a soft material.
  • it further comprises a cover member made of a hard material and overlapping and abutting on the plug member.
  • the cover member includes at least a second flange portion covering the first flange portion, a fourth tubular portion covering an inner peripheral surface of the second tubular portion, and the first bottom portion. a second bottom covering a portion.
  • the port has an extension extending from the first bottom to the one side in the first direction, and the cover member is in contact with the outer peripheral surface of the extension.
  • a fifth tubular portion surrounding the outer peripheral surface is included.
  • the first bottom portion includes a thin portion having a relatively smaller thickness than other portions and having gas permeability.
  • the first bottom portion has a thickness relatively smaller than that of other portions and includes a thin portion having gas permeability
  • the second bottom portion includes A through hole is formed to penetrate through, and the through hole overlaps with the thin portion when viewed in the first direction.
  • the tip of the extending portion is provided with a protruding portion that protrudes outward from the outer peripheral surface and can come into contact with the one side end of the fifth tubular portion in the first direction. is provided.
  • the flow path comprises a first flow path open to the outside and a second flow path open to the inner space of the container body and connected to the ceiling recess.
  • the channel and the second channel are spaced apart from each other in the first direction and overlap when viewed in the first direction.
  • FIG. 2 is an exploded perspective view of the cell culture vessel shown in FIG. 1;
  • FIG. 2 is a plan view of the cell culture vessel shown in FIG. 1;
  • 4 is an enlarged cross-sectional view taken along line IV-IV of FIG. 3;
  • FIG. 4 is an enlarged cross-sectional view taken along line VV of FIG. 3;
  • FIG. 4 is a bottom view of a plug member that constitutes the cell culture vessel according to the first embodiment;
  • FIG. 5 is a cross-sectional view, similar to FIG. 4, showing the state of use of the cell culture vessel according to the first embodiment;
  • FIG. 5 is a cross-sectional view, similar to FIG.
  • FIG. 4 showing the state of use of the cell culture vessel according to the first embodiment
  • FIG. 5 is a cross-sectional view, similar to FIG. 4, showing the state of use of the cell culture vessel according to the first embodiment
  • FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a first modified example of the first embodiment
  • FIG. 11 is a bottom view of a plug member that constitutes a cell culture vessel according to a first modified example of the first embodiment
  • FIG. 11 is a bottom view showing another example of the plug member
  • FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a second modified example of the first embodiment
  • FIG. 5 is a cross-sectional view similar to FIG.
  • FIG. 4 showing a cell culture vessel according to a third modified example of the first embodiment
  • FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a fourth modified example of the first embodiment
  • FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a second embodiment
  • FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a third embodiment
  • FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a fourth embodiment
  • an entity A overlaps an entity B when viewed in a certain direction means, unless otherwise specified, “an entity A overlaps all of an entity B” and “an entity A overlaps an entity B.” "A overlaps part of something B”.
  • FIGS. 1 and 3 to 5 show an assembled state in which the container body 1, the plug member 2, the cover member 3 and the outer lid 4 are combined.
  • FIG. 2 is an exploded perspective view of constituent members of the cell culture vessel A1.
  • the container body 1 is provided with a first cylindrical portion 11 and a container bottom portion 12, and has a container shape having an opening 110 at an upper end 114 (one side end in the first direction) of the first cylindrical portion 11. It is
  • the first tubular portion 11 has a substantially cylindrical shape and extends in the vertical direction (first direction).
  • a male screw 111 is formed on the upper outer peripheral surface of the first tubular portion 11 .
  • the container bottom portion 12 is connected to the vicinity of the lower end of the first tubular portion 11 (near the other side end in the first direction) and closes the inside of the first tubular portion 11 .
  • a slit 112 is formed at an appropriate location on the lower end of the first tubular portion 11 .
  • an extension piece 113 is provided at a suitable position on the lower end of the first tubular portion 11 .
  • the extension piece 113 is a portion that is connected to the first cylindrical portion 11 and extends radially outward.
  • the extension piece 113 is provided at a position different from the slit 112 in the circumferential direction of the first tubular portion 11 .
  • the container body 1 is made of, for example, a translucent or transparent plastic material.
  • plastic material include, but are not limited to, polystyrene, methylpentene, polycarbonate, cycloolefin polymer, cycloolefin copolymer, and other transparent materials.
  • the upper surface of the container bottom 12 is a flat cell culture surface for culturing cells.
  • This cell culture surface (the upper surface of the container bottom portion 12) is appropriately subjected to a surface treatment for improving cell adhesiveness, if necessary.
  • the surface treatment include hydrophilic treatment such as corona discharge treatment and plasma treatment.
  • the plug member 2 is for closing the opening 110 of the container body 1 and sealing the container body 1 .
  • the plug member 2 is, for example, a molded rubber product, and includes a second tubular portion 21, a first flange portion 22, a first bottom portion 23 and a plurality of ports 24, as shown in FIGS. consists of
  • the second cylindrical portion 21 has a substantially cylindrical shape, and contacts and seals the inner peripheral surface of the first cylindrical portion 11 .
  • a pair of annular protrusions 211 are formed on the outer peripheral portion of the second tubular portion 21 at intervals in the vertical direction (first direction).
  • the outer diameter dimension of the annular protrusion 211 in the natural state is slightly larger than the inner diameter dimension of the first cylindrical portion 11, and when the plug member 2 is attached to the container body 1, the second cylindrical portion 21 is It is fitted into the first cylindrical portion 11 while being compressed radially inward. Then, the annular protrusion 211 is in close contact with the inner peripheral surface of the first cylindrical portion 11 to seal it.
  • the second tubular portion 21 may be configured without the annular projection 211 . If the annular projection 211 is not provided, the outer peripheral surface of the second tubular portion 21 surface-contacts with the inner peripheral surface of the first tubular portion 11 for sealing.
  • the first flange portion 22 has a substantially annular shape and extends radially outward from the upper end (one side end in the first direction) of the second cylindrical portion 21 .
  • the first flange portion 22 overlaps with the first tubular portion 11 when viewed in the thickness direction of the first flange portion 22 and closes the upper end of the first tubular portion 11 .
  • the first flange portion 22 has an appropriate thickness and has an appropriate elastic restoring force against a load in the vertical direction (first direction).
  • the thickness of the first flange portion 22 is, for example, about 1 to 3 mm.
  • the first bottom portion 23 extends radially inward from the lower end of the second tubular portion 21 (the other end in the first direction), and closes the inside of the second tubular portion 21 when viewed in the vertical direction.
  • the first bottom portion 23 is plate-shaped and has a ceiling surface 231 .
  • the ceiling surface 231 is a flat surface facing downward (the other side in the first direction).
  • a ceiling concave portion 232 is formed in the first bottom portion 23 .
  • the ceiling recess 232 is a portion recessed upward (one side in the first direction) from the ceiling surface 231 .
  • the ceiling recessed portion 232 is provided near the outer periphery of the first bottom portion 23 . Although the details will be described later, the ceiling recess 232 is arranged corresponding to the port 24 .
  • the first bottom portion 23 has a thin portion 238 .
  • the thin portion 238 is a portion that is thinner than other portions.
  • the thin portion 238 is arranged in the center of the first bottom portion 23 and has a substantially circular shape when viewed in the vertical direction (first direction).
  • the top surface of the first bottom portion 23 is recessed at the central portion thereof, and the recessed portion of the top surface of the first bottom portion 23 is the thin portion 238 .
  • FIG. 6 the boundary between the thin portion 238 and a portion surrounding the thin portion 238 when viewed in the thickness direction (first direction) (hereinafter referred to as "peripheral thick portion 235" is indicated by a virtual line.
  • the thin portion 238 has gas permeability. Also in this embodiment, thinned portion 238 is transparent.
  • the thickness of the thin portion 238 is, for example, approximately 0.2 to 0.3 mm.
  • the thickness of the outer peripheral thick portion 235 surrounding the thin portion 238 is about the same as the thickness of the first flange portion 22, for example, about 1 to 3 mm.
  • a plurality of ports 24 are provided on the first bottom portion 23 and are integrally formed at appropriate locations on the first bottom portion 23 .
  • the first bottom portion 23 is provided with a pair of ports 24 .
  • the pair of ports 24 are provided near the outer periphery of the first bottom portion 23 and are arranged on opposite sides of the center of the first bottom portion 23 when viewed in the vertical direction (first direction).
  • Each port 24 is provided in the outer thick portion 235 .
  • the port 24 is for supplying the culture solution to the inside of the container body 1 or discharging the culture solution inside the container body 1 to the outside, and as shown in FIG. have.
  • the extending portion 241 has a cylindrical shape extending upward (one side in the first direction) from the first bottom portion 23 and has an outer peripheral surface 241a having a circular cross section.
  • a projection 245 is provided at the tip of the extension 241 and projects outward from the outer peripheral surface 241a.
  • the flow path 242 has an upper end (one side end in the first direction) open to the outside and a lower end (hand side end in the first direction) open to the inner space of the container body 1 so that the outside and the container body are open. It is a communication passage leading to the inner space of 1.
  • the channel 242 has a circular cross section and is formed in the extension 241 .
  • the ceiling recessed portion 232 overlaps the port 24 when viewed in the vertical direction (first direction) and is connected to the flow path 242 .
  • the ceiling recessed portion 232 has a conical shape, and is formed such that the area of the cross section increases downward from the position where it is connected to the flow path 242 .
  • the ceiling recess 232 is formed across the first bottom portion 23 and the port 24 (extending portion 241).
  • the plug member 2 configured as described above is made of a soft material having flexibility and elasticity.
  • the material forming the plug member 2 include silicone rubber and elastomer resin.
  • the material of the plug member 2 is more preferably non-cytotoxic and biocompatible medical silicone rubber.
  • the hardness of the plug member 2 it is preferable that the rubber hardness is about 20 to 40 degrees, for example.
  • the cover member 3 overlaps the plug member 2 and contacts the plug member 2 .
  • the cover member 3 includes a second flange portion 31 , a fourth tubular portion 32 , a second bottom portion 33 and a fifth tubular portion 34 .
  • the second flange portion 31 covers the first flange portion 22 from above and is in contact with the upper surface of the first flange portion 22 .
  • the fourth tubular portion 32 is connected to the inner peripheral edge of the second flange portion 31 .
  • the fourth tubular portion 32 has a substantially cylindrical shape and extends downward (the other side in the first direction) from the inner peripheral edge of the second flange portion 31 .
  • the fourth tubular portion 32 covers the inner peripheral surface of the second tubular portion 21 and is in contact with the inner peripheral surface of the second tubular portion 21 .
  • the second bottom portion 33 is connected to the lower end of the fourth tubular portion 32 (the other end in the first direction) and extends radially inward from the lower end of the fourth tubular portion 32 .
  • the second bottom portion 33 covers the first bottom portion 23 from above and is in contact with the upper surface of the first bottom portion 23 .
  • the central portion of the second bottom portion 33 is recessed downward (on the other side in the first direction) corresponding to the location where the thin portion 238 is formed.
  • a through hole 331 is formed in the second bottom portion 33 .
  • the through hole 331 penetrates the second bottom portion 33 in the thickness direction and overlaps the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction).
  • a plurality of through holes 331 are formed in the second bottom portion 33 . As shown in FIG. 3 and the like, the plurality of through holes 331 are appropriately distributed when viewed in the vertical direction.
  • the fifth tubular portion 34 is provided on the second bottom portion 33 and extends upward (one side in the first direction) from a suitable location on the second bottom portion 33 .
  • the second bottom portion 33 is provided with a pair of fifth cylindrical portions 34 .
  • the pair of fifth cylindrical portions 34 are arranged corresponding to the pair of ports 24, respectively.
  • each fifth tubular portion 34 is shaped to be connected to a portion of the fourth tubular portion 32 .
  • the port 24 (extending portion 241) is fitted in each of the pair of fifth cylindrical portions 34.
  • the inner diameter dimension of the fifth tubular portion 34 is slightly smaller than the outer diameter dimension of the extending portion 241 in the natural state.
  • Each extending portion 241 is press-fitted into the fifth tubular portion 34 .
  • the fifth tubular portion 34 is in close contact with (contacts with) the outer peripheral surface 241a of the extending portion 241 to surround the outer peripheral surface 241a.
  • the fifth cylindrical portion 34 overlaps the protrusion 245 provided at the tip of the port 24. there is The projecting portion 245 is adjacent above the fifth tubular portion 34 . As a result, the protrusion 245 can come into contact with the upper end of the fifth tubular portion 34 .
  • the cover member 3 is made of a hard material, such as a translucent or transparent plastic material.
  • a hard material such as a translucent or transparent plastic material.
  • the plastic material include, but are not limited to, polystyrene, methylpentene, polycarbonate, cycloolefin polymer, cycloolefin copolymer, and other transparent materials.
  • the outer lid 4 has a top plate portion 41 and a third tubular portion 42 .
  • the top plate portion 41 has a substantially annular shape and is a portion that sandwiches the second flange portion 31 of the cover member 3 and the upper end 114 of the first cylindrical portion 11 of the container body 1 .
  • a through hole 411 is formed radially inside the top plate portion 41 .
  • the through hole 411 is surrounded by the top plate portion 41 when viewed in the thickness direction of the top plate portion 41 .
  • the through hole 411 overlaps the thin portion 238 of the plug member 2 and the plurality of through holes 331 of the cover member 3 when viewed in the thickness direction (vertical direction) of the top plate portion 41 .
  • the third tubular portion 42 extends from the outer peripheral edge of the top plate portion 41 in the thickness direction (downward in the figure) of the top plate portion 41 and has a substantially cylindrical shape.
  • a female screw 421 is formed on the inner peripheral surface of the third tubular portion 42, and the female screw 421 can be screwed into the male screw 111 of the container body 1 (first tubular portion 11).
  • the top plate portion 41 is provided with a convex portion 412 .
  • the convex portion 412 protrudes upward (one side in the first direction) from the main surface 41a of the top plate portion 41 facing the thickness direction.
  • a plurality of convex portions 412 are provided separately from each other.
  • a plane P ⁇ b>1 formed by the tips 412 a of these convex portions 412 is substantially parallel to the main surface 41 a of the top plate portion 41 .
  • substantially parallel is designed so that the plane P1 and the main surface 41a are parallel, but it also includes the case where there is some variation in parallelism due to dimensional tolerance or the like.
  • a plane P1 formed by the tips 412a of the plurality of convex portions 412 is located at the highest position in the cell culture container A1.
  • the convex portions 412 are provided in the shape of concentric arcs when viewed in the up-down direction, and gaps are formed between adjacent convex portions 412 .
  • the convex portion 412 does not form a closed annular shape when viewed in the vertical direction (thickness direction of the top plate portion 41).
  • the container bottom portion 12 of the container body 1 is connected to a skirt portion 121 projecting downward from the outer peripheral edge thereof.
  • the outer diameter dimension of the outer peripheral edge of the concentric arcs formed by the plurality of convex portions 412 is slightly smaller than the inner diameter dimension of the skirt portion 121 . According to such a configuration, the convex portion 412 of the cell culture vessel A1 can be fitted into the skirt portion 121 of the other cell culture vessel A1 so that the other cell culture vessel A1 can be stacked. Therefore, a plurality of cell culture vessels A1 can be stacked in a stable posture.
  • the outer lid 4 is made of, for example, an opaque, translucent or transparent plastic material.
  • plastic material include, but are not limited to, polyethylene, polypropylene, polystyrene, methylpentene, and polycarbonate.
  • the cell culture vessel A1 is used for perfusion culture in which the culture solution is automatically supplied at a constant low speed and the same amount of culture solution is discharged at the same time. It should be noted that the cultured cells and the culture solution accommodated in the cell culture container A1 (container body 1) are not particularly limited.
  • each port 24 and the connector 7 are assembled in advance (see Fig. 7).
  • the connector 7 is press-fitted to each of the pair of ports 24 .
  • a perfusion pump (not shown) is connected to one port 24 via a connector 7 and a tube 8
  • a drainage container (not shown) is connected to the other port 24 via a connector 7 and a tube 8.
  • Each tube 8 is passed through the inner side of the outer lid 4 in advance.
  • cells to be cultured are seeded in the container body 1, and a predetermined amount of culture solution is placed in the container body 1.
  • FIG. 8 shows a state in which the container body 1 is filled with the culture medium C.
  • the culture solution C fills the inner space of the container body 1 (the space surrounded by the container bottom portion 12 and the first cylindrical portion 11 of the container body 1 and the first bottom portion 23 of the plug member 2).
  • a small amount of air hereinafter referred to as “bubble B” as appropriate may remain in the inner space of the container body 1 .
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is a flat surface.
  • a ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 .
  • the air bubbles B in the container body 1 travel along the ceiling surface 231 and near the port 24. move to By operating the perfusion pump in this state, the air bubbles B remaining in the container body 1 can be further moved into the channel 242 through the ceiling recess 232 and discharged to the outside of the cell culture container A1.
  • a new culture solution C is supplied to the container body 1 (cell culture container A1) through one port 24, and the container body 1 (inside the cell culture container A1) is supplied through the other port 24. of the culture solution C is discharged to the outside.
  • the culture solution C in the vessel body 1 is continuously replaced little by little.
  • the cell culture vessel A1 has an outer lid 4 in addition to the vessel body 1 and the plug member 2.
  • a female screw 421 is formed in the third tubular portion 42 of the outer lid 4 fitted onto the first tubular portion 11 of the container body 1 .
  • the first flange portion 22 of the plug member 2 is connected to the upper end 114 of the first cylindrical portion 11 and the outer lid 4 .
  • the assembled state is maintained by being sandwiched between the top plate portion 41 of the .
  • the plug member 2 is in the form of a series of films extending from the first flange portion 22 on the outer peripheral side to the central first bottom portion 23 to the thin portion 238 .
  • the contents (cultured cells and culture medium C) housed in the container body 1 are liquid-sealed. Therefore, in the perfusion culture process, the culture solution perfusion work can be performed while maintaining the closed state of the cell culture vessel A1, and the risk of contamination can be avoided. Therefore, according to the cell culture vessel A1, it is possible to keep the culture solution C in good condition and ensure the quality of the cells.
  • the male screw 111 of the container body 1 (first tubular portion 11) and the female screw 221 of the outer lid 4 (third tubular portion 42) correspond to an example of the "locking means" of the present disclosure.
  • the cell culture vessel A1 includes a cover member 3 in addition to the container body 1 and the plug member 2.
  • the plug member 2 is made of a soft material.
  • the cover member 3 is made of a hard material and is in contact with the plug member 2 in an overlapping manner.
  • the cover member 3 includes a second flange portion 31 that covers the first flange portion 22 of the plug member 2 .
  • the second flange portion 31 is interposed between the first flange portion 22 of the plug member 2 and the top plate portion 41 of the outer lid 4 . Therefore, when the female screw 421 of the outer lid 4 is screwed into the male screw 111 of the container body 1 (first cylindrical portion 11), the top plate portion 41 is in sliding contact with the second flange portion 31 (hard material). slidability becomes good.
  • the cover member 3 includes a fourth tubular portion 32 that covers the inner peripheral surface of the second tubular portion 21 . According to such a configuration, the plug member 2 made of a soft material is pressed against the inner peripheral surface of the container body 1 (first tubular portion 11), and the sealing performance of the cell culture container A1 is enhanced.
  • the cover member 3 also includes a second bottom portion 33 that covers the first bottom portion 23 of the plug member 2 . According to such a configuration, in the perfusion culture process, the first bottom portion 23 (thin portion 238) tries to swell upward due to the pressurizing force of the supplied culture medium C, but the second bottom portion 33 causes the first bottom portion 23 to expand. Deformation of the bottom portion 23 (thin portion 238) is suppressed. As a result, the flow of the culture solution C inside the container body 1 becomes smooth, and the visibility inside the container body 1 is excellent, making it suitable for observing the conditions inside the container body 1 .
  • the port 24 provided on the plug member 2 has an extending portion 241 extending upward from the first bottom portion 23 .
  • the cover member 3 includes a fifth cylindrical portion 34, and the fifth cylindrical portion 34 contacts the outer peripheral surface 241a of the extending portion 241 to cover the outer peripheral surface 241a.
  • the first bottom portion 23 of the plug member 2 includes a thin portion 238 having gas permeability. Therefore, the contents of the container body 1 (cell culture container A1) are maintained in a ventilated state with the outside of the container body 1 (cell culture container A1). Therefore, according to the present embodiment, the contents of the container body 1 (cell culture container A1) can be cultured in an aerated state with the outside. Further, if the cell culture vessel A1 is placed in an incubator under a predetermined gas atmosphere, the gas atmosphere in the incubator can be taken into the container body 1 (cell culture vessel A1).
  • a through hole 331 is formed in the second bottom portion 33 of the cover member 3 so as to pass through the second bottom portion 33 in the thickness direction.
  • the through hole 331 overlaps the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction). According to such a configuration, while suppressing the deformation of the first bottom portion 23 (the thin portion 238 ) as described above, it is possible to maintain the state of ventilation between the inside and the outside of the container body 1 via the thin portion 238 . .
  • a protrusion 245 that protrudes outward from the outer peripheral surface 241 a of the extension 241 is provided at the tip of the port 24 (extension 241 ).
  • the projecting portion 245 is adjacent to the upper side of the fifth tubular portion 34 and can abut on the upper end of the fifth tubular portion 34 . According to such a configuration, even if a downward pressing force acts on the extending portion 241, the extending portion 241 is prevented from being pushed into the inner space of the container body 1 through the fifth cylindrical portion 34. .
  • a downward pressing force acts on the port 24, but the extending portion 241 is prevented from entering the inner space of the container body 1 unreasonably. Damage to cells in 1 is avoided.
  • FIG. 10 shows a cell culture vessel according to a first modified example of the first embodiment.
  • the same or similar elements as those of the cell culture vessel A1 of the above embodiment are denoted by the same reference numerals as those of the above embodiment, and description thereof will be omitted as appropriate.
  • the cell culture vessel A11 of this modified example differs from the cell culture vessel A1 of the above embodiment mainly in the configuration of the ceiling recess 232 .
  • the ceiling concave portion 232 has a conical portion 233 and a peripheral edge portion 234 .
  • the conical portion 233 is connected to the channel 242 .
  • the cone-shaped portion 233 is a cone-shaped portion formed so that the cross-sectional area increases downward from the position where it connects with the flow path 242 .
  • the peripheral edge portion 234 connects to the conical portion 233 and communicates with the outer peripheral edge 237 of the first bottom portion 23 .
  • the peripheral edge portion 234 is inclined downward as it goes radially outward of the first bottom portion 23 . As shown in FIG.
  • the peripheral edge portion 234 has a substantially fan shape extending radially outward when viewed in the thickness direction (first direction) of the first bottom portion 23 .
  • the annular protrusion 211 is not formed on the outer peripheral portion of the second tubular portion 21 , and the outer peripheral surface of the second tubular portion 21 faces the inner peripheral surface of the first tubular portion 11 . are in direct contact.
  • the cell culture vessel A11 of this modified example is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time.
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat.
  • a ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position.
  • the ceiling recessed portion 232 (peripheral edge portion 234 ) communicates with the outer peripheral edge 237 of the first bottom portion 23 .
  • the cell culture vessel A11 of this modified example has the same effects as the cell culture vessel A1 described above.
  • FIG. 12 is a bottom view showing another example of the plug member 2.
  • FIG. The plug member 2 shown in FIG. 12 differs from the plug member 2 shown in FIG. 11 in the shape of the peripheral portion 234 .
  • the peripheral edge portion 234 is formed to extend along the circumferential direction on the outer peripheral portion of the first bottom portion 23 .
  • the peripheral edge portion 234 is inclined so as to be positioned downward as it goes radially outward of the first bottom portion 23 .
  • the air bubbles can be guided to the flow path 242 along the peripheral edge portion 234 . Thereby, it is possible to more accurately discharge the air bubbles remaining in the inner space of the container body 1 .
  • FIG. 13 shows a cell culture vessel according to a second modification of the first embodiment.
  • the configurations of the first bottom portion 23 of the plug member 2 and the second bottom portion 33 of the cover member 3 are different from those of the cell culture vessel A1 of the above embodiment.
  • the first bottom portion 23 (mainly the thin-walled portion 238) has a curved plate shape, and curves downward (on the other side in the first direction) as it extends radially inward.
  • the ceiling surface 231 is a curved surface that curves downward as it extends radially inward.
  • the second bottom portion 33 is also curved so as to be positioned downward (on the other side in the first direction) as it goes radially inward.
  • the portion of the second bottom portion 33 that overlaps the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction) is flat (see FIG. 4).
  • the second bottom portion 33 has a curved portion overlapping the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction).
  • the lower surface of the second bottom portion 33 is in contact with the thin portion 238 .
  • the thin portion 238 may have a curved shape in its natural state, but is not limited to this.
  • the thin portion 238 may be flat in its natural state, and may be curved when covered with the second bottom portion 33 of the cover member 3 .
  • the cell culture container A12 of this modified example is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time.
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is curved.
  • a ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the air bubbles remain on the curved ceiling surface 231.
  • the cell culture vessel A12 of this modified example has the same effects as the cell culture vessel A1 described above.
  • FIG. 14 shows a cell culture vessel according to a third modified example of the first embodiment.
  • the configuration of one of the pair of ports 24 is different from that of the cell culture vessel A1 of the above embodiment.
  • the port 24 located on the left side of FIG. 14 is the same as that of the above embodiment, but the configuration of the port 24 located on the right side of the figure is different.
  • the port 24 located on the right side of FIG. 14 has a protrusion 247 extending below the first bottom 23 .
  • the ceiling recess 232 is not formed around the projecting portion 247 in the first bottom portion 23 .
  • the cell culture vessel A13 of this modified example is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time.
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat.
  • the first bottom portion 23 is formed with a ceiling recess 232 recessed upward from the ceiling surface 231 , and the ceiling recess 232 is connected to a flow path 242 of one port 24 (left side in FIG. 14).
  • one port 24 (left side in FIG. 14) ) is positioned at the top, air bubbles in the container body 1 move along the ceiling surface 231 to the vicinity of the one port 24 . Then, the air bubbles float in the channel 242 through the ceiling recess 232 . Thereby, the air bubbles remaining in the container body 1 can be discharged to the outside of the cell culture container A13. Then, when perfusion culture is performed, a new culture solution C is supplied to the container body 1 (cell culture container A13) through the port 24 located on the right side of FIG. The culture solution inside the body 1 (inside the cell culture container A13) is discharged to the outside. In the perfusion culture process, the culture solution in the container body 1 is continuously replaced little by little.
  • the cell culture vessel A13 of this modified example has the same effects as the cell culture vessel A1 described above.
  • FIG. 15 shows a cell culture vessel according to a fourth modified example of the first embodiment.
  • the configuration of the second bottom portion 33 of the cover member 3 is different from that of the cell culture vessel A1 of the above embodiment.
  • one through hole 331 is formed in the second bottom portion 33 .
  • the diameter of the through hole 331 is remarkably increased as compared with the above embodiment.
  • the through hole 331 overlaps most of the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction).
  • the cell culture vessel A14 of this modification is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time.
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat.
  • a ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position.
  • the cell culture vessel A14 of this modified example has the same effects as the cell culture vessel A1 described above.
  • FIG. 16 shows a cell culture vessel according to a second embodiment of the present disclosure.
  • the cell culture vessel A2 of this embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the port 24 .
  • the port 24 has a first channel 243, a second channel 244, and a non-penetrating portion 246 instead of the channel 242 of the above embodiment.
  • the flow path 242 in the first embodiment is a communication path that communicates with the outside and the inner space of the container body 1, but in the present embodiment, the first flow path 243 and the second flow path 244 are , are separated from each other in the vertical direction (first direction).
  • the first channel 243 is located near the upper portion of the port 24 and is open to the outside.
  • the second channel 244 is located near the bottom of the port 24 , opens to the inner space of the container body 1 , and is connected to the ceiling recess 232 .
  • the first flow path 243 and the second flow path 244 overlap each other when viewed in the vertical direction (first direction).
  • the non-penetrating portion 246 is a portion interposed between the first channel 243 and the second channel 244 .
  • the cell culture vessel A2 of the present embodiment is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of the culture solution is discharged at the same time.
  • the introduction of cells and culture solution into the inner space of the container body 1 and the discharge of the waste liquid are performed, for example, by attaching the plug member 2, the cover member 3 and the outer lid 4 to the container body 1, and After A1 is brought into an assembled state, the non-penetration portion 246 of the port 24 is pierced with an injection needle (not shown).
  • the tip of the injection needle is arranged in the second flow path 244 immediately below the non-penetrating portion 246 .
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is a flat surface.
  • a ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 .
  • the port 24 provided on the plug member 2 has an extending portion 241 extending upward from the first bottom portion 23 .
  • the cover member 3 includes a fifth cylindrical portion 34, and the fifth cylindrical portion 34 contacts the outer peripheral surface 241a of the extending portion 241 to cover the outer peripheral surface 241a.
  • the cell culture vessel A2 of this embodiment has the same effects as the cell culture vessel A1 described above.
  • FIG. 17 shows a cell culture vessel according to the third embodiment of the present disclosure.
  • the cell culture vessel A3 of this embodiment does not include the cover member 3 unlike the cell culture vessel A1 of the first embodiment. That is, the cell culture vessel A3 has a vessel body 1, a plug member 2 and an outer lid 4. As shown in FIG.
  • the configurations of the vessel body 1 and the outer lid 4 are the same as in the first embodiment.
  • the configuration of the plug member 2 is significantly different from that of the first embodiment.
  • the plug member 2 is configured with a first portion 2A and a second portion 2B.
  • the second part 2B is made of a soft material having flexibility and elasticity, and has a second tubular part 21 and a first flange part 22 .
  • the second portion 2B is interposed between the first portion 2A and the first cylindrical portion 11 of the container body 1.
  • a pair of annular projections 211 are formed on the outer peripheral portion of the second cylindrical portion 21 with a gap therebetween in the vertical direction (first direction).
  • the first portion 2A includes a first bottom portion 23, a port 24, an inner cylindrical portion 26 and an upper flange portion 27.
  • the first bottom portion 23 has a substantially constant thickness. Accordingly, in this embodiment, unlike the first embodiment, the first bottom portion 23 does not have the thin portion 238 .
  • a ceiling recess 232 formed in the first bottom portion 23 has a conical portion 233 and a peripheral portion 234 .
  • the conical portion 233 is connected to the channel 242 .
  • the cone-shaped portion 233 is a cone-shaped portion formed so that the cross-sectional area increases downward from the position where it connects with the flow path 242 .
  • the peripheral edge portion 234 is connected to the conical portion 233 and communicates with the outer peripheral edge of the first bottom portion 23 .
  • the peripheral edge portion 234 is inclined downward as it goes radially outward of the first bottom portion 23 .
  • the inner tubular portion 26 extends upward from the outer peripheral edge of the first bottom portion 23 and has a substantially cylindrical shape.
  • the inner tubular portion 26 covers the inner peripheral surface of the second tubular portion 21 .
  • the upper flange portion 27 has a substantially annular shape and extends radially outward from the upper end of the inner cylindrical portion 26 .
  • the upper flange portion 27 overlaps the first flange portion 22 and the first tubular portion 11 when viewed in the thickness direction of the upper flange portion 27 .
  • the first part 2A is made of a hard material, such as a translucent or transparent plastic material.
  • a hard material such as a translucent or transparent plastic material.
  • the plastic material include, but are not limited to, polystyrene, methylpentene, polycarbonate, cycloolefin polymer, cycloolefin copolymer, and other transparent materials.
  • the cell culture vessel A3 of the present embodiment is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of the culture solution is discharged at the same time.
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat.
  • a ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position.
  • the ceiling recessed portion 232 (peripheral edge portion 234) communicates with the outer peripheral edge of the first bottom portion 23.
  • the air bubbles are removed from the outer peripheral edge of the first bottom portion 23 or the outer peripheral edge of the second cylindrical portion 21. Even if it reaches the edge side, the air bubble can be guided to the channel 242 along the peripheral edge portion 234 . Thereby, it is possible to more accurately discharge the air bubbles remaining in the inner space of the container body 1 .
  • the cell culture vessel A3 of the present embodiment has the same structure as the cell culture vessel A1 of the first embodiment described with reference to FIGS. It has the same function and effect as.
  • FIG. 18 shows a cell culture vessel according to a fourth embodiment of the present disclosure.
  • the cell culture vessel A4 of this embodiment does not have the cover member 3 and the outer lid 4.
  • the cell culture vessel A4 has a container body 1 and a plug member 2.
  • FIG. 18 shows a container body 1 and a plug member 2.
  • the external thread 111 is not formed on the outer peripheral surface of the first cylindrical portion 11 of the container body 1 .
  • the plug member 2 is, for example, a molded rubber product, and includes a second cylindrical portion 21, a first flange portion 22, a first bottom portion 23 and a pair of ports 24.
  • the plug member 2 is made of a soft material having flexibility and elasticity.
  • the material constituting the plug member 2 is the same as that of the first embodiment.
  • the annular protrusion 211 is not formed on the outer peripheral portion of the second tubular portion 21 .
  • the outer diameter dimension of the second tubular portion 21 is larger than the inner diameter dimension of the first tubular portion 11 in the natural state.
  • the second tubular portion 21 configured as described above is press-fitted into the first tubular portion 11 .
  • the outer peripheral surface of the second cylindrical portion 21 is in close contact with (abuts) the inner peripheral surface of the first cylindrical portion 11 to form a tight seal.
  • the first bottom portion 23 does not have the thin portion 238 .
  • the ceiling concave portion 232 formed in the first bottom portion 23 has a conical shape and is formed across the port 24 (extending portion 241) from the first bottom portion 23. As shown in FIG.
  • the cell culture vessel A4 of this embodiment is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of the culture solution is discharged at the same time.
  • the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat.
  • a ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position.
  • the cell culture vessel A4 of the present embodiment has the same structure as the cell culture vessel A1 of the first embodiment described with reference to FIGS. It has the same function and effect as.
  • the cell culture vessel according to the present disclosure is not limited to the above embodiments.
  • the specific configuration of each part of the cell culture vessel of the present disclosure can be changed in various ways.
  • A1, A11, A12, A13, A14, A2, A3, A4 cell culture vessel
  • B air bubble
  • C culture solution
  • 1 container body
  • 11 first cylindrical portion
  • 110 opening
  • 111 male screw (locking means)
  • 112 slit
  • 113 extension piece
  • 114 upper end (first direction one side end of the first cylindrical portion)
  • 12 container bottom
  • 121 skirt portion
  • 2 plug member
  • 2A first part
  • 2B second part
  • 21 second tubular part
  • 232 ceiling concave part
  • 233 cone shape portion 234: peripheral portion 235: outer peripheral side thick portion 237: outer peripheral edge 238: thin portion
  • 241: extension portion 241a outer peripheral surface
  • 242 flow path 243: first Flow path 244: Second flow path 245: Protruding portion 246: Non-penetrating portion 247: Protruding portion 26: Inner cylindrical

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Abstract

Provided is a cell culture vessel suited to improving difficulties when performing perfusion culture in a closed state. The cell culture vessel is equipped with a vessel body 1 having an opening 110 on the upper end and a first cylindrical portion 11 extending vertically, and a plug member 2 capable of sealing the opening 110. The plug member 2 includes: a second cylindrical portion 21 that abuts the inner peripheral surface and closes the first cylindrical portion 11; a first flange portion 22 that extends radially outward from the upper end of the second cylindrical portion 21 and blocks the upper end 114 of the first cylindrical portion 11; and a first bottom portion 23 that extends radially inward from the lower end of the second cylindrical portion 21 and blocks the inside of the second cylindrical portion 21 when viewed in the vertical direction. The first bottom portion 23 is provided with a plurality of ports 24 each having a flow path 242 leading to the outside and the inside space of the vessel body 1. The first bottom portion 23 has a ceiling surface 231 that faces downward, and ceiling recesses 232 recessed upward from the ceiling surface 231 are formed in the first bottom portion 23. The ceiling recesses 232 connect to the flow paths 242.

Description

細胞培養容器cell culture vessel
　本開示は、細胞を培養するための細胞培養容器に関し、特に、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に用いる細胞培養容器に関する。 The present disclosure relates to a cell culture vessel for culturing cells, and in particular, to a cell culture vessel used for perfusion culture that automatically supplies culture medium at a constant low speed and discharges the same amount of culture medium at the same time.
　細胞の培養に際しては、例えばディッシュ（シャーレ）、ウェルプレート、フラスコなどの培養容器が広く用いられている。ディッシュやウェルプレートは蓋があっても載せるだけで隙間があり、密閉できなかった。フラスコの場合は密閉可能だが、これら従来の培養容器はいずれも培養液の交換等の作業に蓋の開閉が必要であった。そのため、これらの培養容器は、研究を目的にした細胞培養用途には汎用性が高いが、医療を目的にした細胞培養用途には、コンタミネーションのリスク、専門技術を持つ人材やコストの問題、培養細胞の品質均一化が難しいという問題があった。 For culturing cells, culture vessels such as dishes (petri dishes), well plates, and flasks are widely used. Even if the dishes and well plates had lids, there was a gap just by putting them on, and it was impossible to seal them. Flasks can be hermetically sealed, but all of these conventional culture vessels require the opening and closing of lids for operations such as exchanging the culture solution. Therefore, these culture vessels are highly versatile for use in cell culture for research purposes. There was a problem that it was difficult to equalize the quality of cultured cells.
　昨今、これらの問題を解決するべく様々な自動培養装置が開発されている（例えば、特許文献１を参照）。自動培養装置に用いられる培養容器において、例えば一対のポートが設けられている。これらポートは、例えば培養容器の蓋に設けられ、それぞれ蓋から下方に延びて培養容器の内部に配置される。細胞培養を行う際には、一方のポートを通じて新しい培養液が培養容器に供給され、かつ他方のポートを通じて培養容器内部の培養液が外部に排出される。一方のポートから培養液を供給し他方のポートから排出をするには、それぞれのポートに適したポンプを接続すればよく、その場合容器内に培養液と気体の層があっても液は流れる。しかしながら、一方のポートから供給する培養液と他方のポートから排出する培養液の量が正確に一致しなければ、培養が長期間に及んだ場合には容器内の液が枯渇または溢れることになる。 Recently, various automatic culture devices have been developed to solve these problems (see Patent Document 1, for example). A culture vessel used in an automatic culture apparatus is provided with, for example, a pair of ports. These ports are provided, for example, in the lid of the culture vessel and extend downward from the lid to be arranged inside the culture vessel. When culturing cells, a new culture solution is supplied to the culture container through one port, and the culture solution inside the culture container is discharged to the outside through the other port. In order to supply the culture medium from one port and discharge it from the other port, it is sufficient to connect a suitable pump to each port, in which case the liquid will flow even if there are layers of culture medium and gas in the container . However, if the amount of culture medium supplied from one port and the amount of culture medium discharged from the other port are not exactly the same, the liquid in the container may dry up or overflow during long-term cultivation. Become.
　このような不都合を回避するために、１つの容器に対してポンプを１つにした上で、一方のポートから陽圧で培養液を供給し他方のポートからオーバーフローさせて排出するか、片側のポートから培養液を陰圧で排出しもう一方のポートから培養液を引き込む方法が有効になる。これを一気に行うと乱流が起こり細胞にダメージを与えるため、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養が行われる。そして、この灌流培養においては、スムーズな液流を実現するために、閉鎖系であることと、培養容器の内部全体に培養液を満たし気体の層や気泡を作らないことが重要となる。しかしながら、閉鎖系の培養容器内部を培養液で満たそうとしても、容器内部に少量の空気（気泡）が残存する場合がある。上記のようにポートが蓋から下方に延びて容器内部に配置されていると、容器内部に残存する空気を外部に排出することができなかった。 In order to avoid such inconvenience, one pump is used for one container, and the culture solution is supplied from one port under positive pressure and discharged from the other port by overflowing, or A method in which the culture medium is discharged from the port under negative pressure and the culture medium is drawn in from the other port is effective. If this is done all at once, turbulence will occur and the cells will be damaged. Therefore, perfusion culture is performed in which the culture solution is automatically supplied at a constant low speed and the same amount of culture solution is discharged at the same time. In this perfusion culture, in order to realize smooth liquid flow, it is important to have a closed system and to fill the entire interior of the culture vessel with the culture medium without creating a gas layer or air bubbles. However, even if an attempt is made to fill the inside of a closed-system culture vessel with a culture solution, a small amount of air (bubbles) may remain inside the vessel. When the port extends downward from the lid and is arranged inside the container as described above, the air remaining inside the container cannot be discharged to the outside.
特開２０１７－７９６３３号公報JP 2017-79633 A
　本開示は、このような事情のもとで考え出されたものであって、閉鎖系で灌流培養を行う場合の不都合を改善するのに適した細胞培養容器を提供することを主たる課題とする。 The present disclosure has been conceived under such circumstances, and a main object of the present disclosure is to provide a cell culture vessel suitable for improving the inconvenience of performing perfusion culture in a closed system. .
　上記の課題を解決するため、本開示では、次の技術的手段を採用した。 In order to solve the above problems, the following technical measures are adopted in this disclosure.
　本開示によって提供される細胞培養容器は、第１方向の一方側端に開口を有し、かつ前記第１方向に延びる第１筒状部を有する容器体と、前記開口を閉塞可能な栓部材と、を備え、前記栓部材は、前記第１筒状部の内周面に当接して密閉する第２筒状部、前記第２筒状部の前記第１方向における前記一方側端から径方向外方に延出し、前記第１筒状部の前記第１方向における前記一方側端を塞ぐ第１フランジ部、および前記第２筒状部の前記第１方向における他方側端から径方向内方に延出し、前記第１方向に見て前記第２筒状部の内側を塞ぐ板状の第１底部、を含み、前記第１底部には、前記第１方向の前記一方側端が外部に開放し、かつ前記第１方向の前記他方側端が前記容器体の内側空間において開放する流路を各々が有する複数のポートが設けられており、前記第１底部は、前記第１方向の他方側を向く天井面を有し、前記第１底部には、前記天井面から前記第１方向の前記一方側に凹む天井凹部が形成されており、前記天井凹部は、前記複数のポートのうち少なくともいずれか１つにおける前記流路につながっている。 A cell culture vessel provided by the present disclosure includes a vessel body having an opening at one end in a first direction and having a first cylindrical portion extending in the first direction, and a plug member capable of closing the opening. and, the plug member includes a second cylindrical portion that abuts and seals the inner peripheral surface of the first cylindrical portion, and a diameter from the one side end of the second cylindrical portion in the first direction. a first flange portion that extends outward in the direction and closes the one side end of the first tubular portion in the first direction; a plate-like first bottom part that extends in the direction of the first direction and closes the inside of the second cylindrical part when viewed in the first direction, and the first bottom part has the one side end in the first direction that extends to the outside. a plurality of ports each having a flow path that opens to the first direction and that the other side end in the first direction opens in the inner space of the container body, and the first bottom portion is provided in the first direction a ceiling surface facing the other side, a ceiling recess recessed from the ceiling surface toward the one side in the first direction is formed in the first bottom portion, and the ceiling recess is one of the plurality of ports; Communicate with at least one of the channels.
　好ましい実施の形態においては、前記天井凹部は、前記第１底部の外周縁に通じる。 In a preferred embodiment, the ceiling recess communicates with the outer peripheral edge of the first bottom.
　好ましい実施の形態においては、環状の頂板部と、この頂板部の外周縁から当該頂板部の厚さ方向に延びており、前記第１筒状部に外嵌される第３筒状部と、を有する外蓋をさらに備え、前記容器体および前記外蓋の少なくともいずれか一方には、前記第１筒状部に前記第１フランジ部が当接する状態で前記第１筒状部と前記第３筒状部との相対移動を阻止する係止手段が設けられている。 In a preferred embodiment, an annular top plate portion, a third tubular portion extending from the outer peripheral edge of the top plate portion in the thickness direction of the top plate portion and fitted onto the first tubular portion, At least one of the container body and the outer lid has the first tubular portion and the third flange portion in contact with the first tubular portion. Locking means are provided to prevent relative movement with the tubular portion.
　好ましい実施の形態においては、前記栓部材は、軟質素材からなる。 In a preferred embodiment, the plug member is made of a soft material.
　好ましい実施の形態においては、硬質素材からなり、前記栓部材に重なって当接するカバー部材をさらに備える。 In a preferred embodiment, it further comprises a cover member made of a hard material and overlapping and abutting on the plug member.
　好ましい実施の形態においては、前記カバー部材は、前記第１フランジ部を覆う第２フランジ部と、前記第２筒状部の内周面を覆う第４筒状部と、前記第１底部の少なくとも一部を覆う第２底部と、を含む。 In a preferred embodiment, the cover member includes at least a second flange portion covering the first flange portion, a fourth tubular portion covering an inner peripheral surface of the second tubular portion, and the first bottom portion. a second bottom covering a portion.
　好ましい実施の形態においては、前記ポートは、前記第１底部から前記第１方向の前記一方側に延びる延出部を有し、前記カバー部材は、前記延出部の外周面に当接して当該外周面を囲う第５筒状部を含む。 In a preferred embodiment, the port has an extension extending from the first bottom to the one side in the first direction, and the cover member is in contact with the outer peripheral surface of the extension. A fifth tubular portion surrounding the outer peripheral surface is included.
　好ましい実施の形態においては、前記第１底部は、他の部位よりも相対的に厚さが小とされ、かつガス透過性を有する薄肉部を含む。 In a preferred embodiment, the first bottom portion includes a thin portion having a relatively smaller thickness than other portions and having gas permeability.
　好ましい実施の形態においては、前記第１底部は、他の部位よりも相対的に厚さが小とされ、かつガス透過性を有する薄肉部を含み、前記第２底部には、前記第１方向に貫通する貫通孔が形成されており、前記貫通孔は、前記第１方向に見て前記薄肉部と重なっている。 In a preferred embodiment, the first bottom portion has a thickness relatively smaller than that of other portions and includes a thin portion having gas permeability, and the second bottom portion includes A through hole is formed to penetrate through, and the through hole overlaps with the thin portion when viewed in the first direction.
　好ましい実施の形態においては、前記延出部の先端には、前記外周面よりも外向きに突出し、かつ前記第５筒状部の前記第１方向における前記一方側端に当接し得る突起部が設けられている。 In a preferred embodiment, the tip of the extending portion is provided with a protruding portion that protrudes outward from the outer peripheral surface and can come into contact with the one side end of the fifth tubular portion in the first direction. is provided.
　好ましい実施の形態においては、前記流路は、外部に開放する第１流路、および前記容器体の前記内側空間に開放し、かつ前記天井凹部がつながる第２流路からなり、前記第１流路および前記第２流路は、前記第１方向に互いに離間し、かつ前記第１方向に見て重なっている。 In a preferred embodiment, the flow path comprises a first flow path open to the outside and a second flow path open to the inner space of the container body and connected to the ceiling recess. The channel and the second channel are spaced apart from each other in the first direction and overlap when viewed in the first direction.
　本開示による細胞培養容器のその他の特徴および利点は、添付図面を参照して以下に行う詳細な説明によって、より明らかとなろう。 Other features and advantages of the cell culture vessel according to the present disclosure will become clearer from the detailed description given below with reference to the accompanying drawings.
第１実施形態に係る細胞培養容器を示す斜視図である。It is a perspective view showing a cell culture container concerning a 1st embodiment. 図１に示す細胞培養容器の分解斜視図である。FIG. 2 is an exploded perspective view of the cell culture vessel shown in FIG. 1; 図１に示す細胞培養容器の平面図である。FIG. 2 is a plan view of the cell culture vessel shown in FIG. 1; 図３のＩＶ－ＩＶ線に沿う拡大断面図である。4 is an enlarged cross-sectional view taken along line IV-IV of FIG. 3; FIG. 図３のＶ－Ｖ線に沿う拡大断面図である。4 is an enlarged cross-sectional view taken along line VV of FIG. 3; FIG. 第１実施形態に係る細胞培養容器を構成する栓部材の底面図である。FIG. 4 is a bottom view of a plug member that constitutes the cell culture vessel according to the first embodiment; 第１実施形態に係る細胞培養容器の使用状態を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view, similar to FIG. 4, showing the state of use of the cell culture vessel according to the first embodiment; 第１実施形態に係る細胞培養容器の使用状態を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view, similar to FIG. 4, showing the state of use of the cell culture vessel according to the first embodiment; 第１実施形態に係る細胞培養容器の使用状態を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view, similar to FIG. 4, showing the state of use of the cell culture vessel according to the first embodiment; 第１実施形態の第１変形例に係る細胞培養容器を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a first modified example of the first embodiment; 第１実施形態の第１変形例に係る細胞培養容器を構成する栓部材の底面図である。FIG. 11 is a bottom view of a plug member that constitutes a cell culture vessel according to a first modified example of the first embodiment; 栓部材の他の例を示す底面図である。FIG. 11 is a bottom view showing another example of the plug member; 第１実施形態の第２変形例に係る細胞培養容器を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a second modified example of the first embodiment; 第１実施形態の第３変形例に係る細胞培養容器を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a third modified example of the first embodiment; 第１実施形態の第４変形例に係る細胞培養容器を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a fourth modified example of the first embodiment; 第２実施形態に係る細胞培養容器を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a second embodiment; 第３実施形態に係る細胞培養容器を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a third embodiment; 第４実施形態に係る細胞培養容器を示す、図４と同様の断面図である。FIG. 5 is a cross-sectional view similar to FIG. 4 showing a cell culture vessel according to a fourth embodiment;
　以下、本開示の好ましい実施の形態を図面を参照しつつ具体的に説明する。 Hereinafter, preferred embodiments of the present disclosure will be specifically described with reference to the drawings.
　本開示において、「ある物Ａがある物Ｂにある方向に見て重なる」とは、特段の断りのない限り、「ある物Ａがある物Ｂのすべてに重なること」、および、「ある物Ａがある物Ｂの一部に重なること」を含む。 In the present disclosure, “an entity A overlaps an entity B when viewed in a certain direction” means, unless otherwise specified, “an entity A overlaps all of an entity B” and “an entity A overlaps an entity B.” "A overlaps part of something B".
　本開示における「第１」、「第２」、「第３」等の用語は、単にラベルとして用いたものであり、必ずしもそれらの対象物に順列を付することを意図していない。 The terms "first", "second", "third", etc. in the present disclosure are merely used as labels and are not necessarily intended to give permutations to those objects.
＜第１実施形態＞
　図１～図５は、本開示の第１実施形態に係る細胞培養容器を示している。本実施形態の細胞培養容器Ａ１は、容器体１、栓部材２、カバー部材３および外蓋４を備えている。詳細は後述するが、図１、図３～図５は、容器体１、栓部材２、カバー部材３および外蓋４を組み合わせたアセンブリ状態を示す。図２は、細胞培養容器Ａ１の構成部材の分解斜視図である。 <First Embodiment>
1 to 5 show a cell culture vessel according to a first embodiment of the present disclosure. A cell culture vessel A1 of this embodiment includes a vessel body 1, a plug member 2, a cover member 3 and an outer lid 4. As shown in FIG. Although the details will be described later, FIGS. 1 and 3 to 5 show an assembled state in which the container body 1, the plug member 2, the cover member 3 and the outer lid 4 are combined. FIG. 2 is an exploded perspective view of constituent members of the cell culture vessel A1.
　本実施形態においては、容器体１は、第１筒状部１１および容器底部１２を備え、第１筒状部１１の上端１１４（第１方向の一方側端）に開口１１０を有する容器状とされている。第１筒状部１１は、概略円筒状とされており、上下方向（第１方向）に延びている。第１筒状部１１の上部外周面には、雄ネジ１１１が形成されている。容器底部１２は、第１筒状部１１の下端近傍（第１方向の他方側端の近傍）につながり、第１筒状部１１の内側を塞いでいる。本実施形態において、第１筒状部１１の下端の適所には、スリット１１２が形成されている。また、本実施形態において、第１筒状部１１の下端の適所には、延出片１１３が設けられている。延出片１１３は、第１筒状部１１につながって径方向外方に延びる部分である。延出片１１３は、第１筒状部１１の周方向においてスリット１１２とは異なる位置に設けられている。 In this embodiment, the container body 1 is provided with a first cylindrical portion 11 and a container bottom portion 12, and has a container shape having an opening 110 at an upper end 114 (one side end in the first direction) of the first cylindrical portion 11. It is The first tubular portion 11 has a substantially cylindrical shape and extends in the vertical direction (first direction). A male screw 111 is formed on the upper outer peripheral surface of the first tubular portion 11 . The container bottom portion 12 is connected to the vicinity of the lower end of the first tubular portion 11 (near the other side end in the first direction) and closes the inside of the first tubular portion 11 . In this embodiment, a slit 112 is formed at an appropriate location on the lower end of the first tubular portion 11 . Further, in the present embodiment, an extension piece 113 is provided at a suitable position on the lower end of the first tubular portion 11 . The extension piece 113 is a portion that is connected to the first cylindrical portion 11 and extends radially outward. The extension piece 113 is provided at a position different from the slit 112 in the circumferential direction of the first tubular portion 11 .
　容器体１は、例えば半透明または透明のプラスチック材料により形成されている。当該プラスチック材料としては、例えばポリスチレンやメチルペンテンのほか、ポリカーボネート、シクロオレフィンポリマー、シクロオレフィンコポリマーなどの、好適には透明性を有する材料が用いられるが、これらに限定されない。 The container body 1 is made of, for example, a translucent or transparent plastic material. Examples of the plastic material include, but are not limited to, polystyrene, methylpentene, polycarbonate, cycloolefin polymer, cycloolefin copolymer, and other transparent materials.
　なお、容器底部１２の上面は、細胞を培養するための平坦な細胞培養面である。この細胞培養面（容器底部１２の上面）には、必要に応じて細胞接着性を向上させるための表面処理が適宜施される。当該表面処理としては、例えばコロナ放電処理、プラズマ処理などの親水化処理を挙げることができる。 The upper surface of the container bottom 12 is a flat cell culture surface for culturing cells. This cell culture surface (the upper surface of the container bottom portion 12) is appropriately subjected to a surface treatment for improving cell adhesiveness, if necessary. Examples of the surface treatment include hydrophilic treatment such as corona discharge treatment and plasma treatment.
　栓部材２は、容器体１の開口１１０を閉塞し、容器体１を密閉するためのものである。栓部材２は、例えばゴム成形品であり、図２、図４、図５等に示すように、第２筒状部２１、第１フランジ部２２、第１底部２３および複数のポート２４を含んで構成される。第２筒状部２１は、概略円筒状とされており、第１筒状部１１の内周面に当接して密閉する。本実施形態では、第２筒状部２１の外周部には一対の環状突起２１１が上下方向（第１方向）に間隔を隔てて形成されている。例えば、自然状態における環状突起２１１の外径寸法は第１筒状部１１の内径寸法よりも少し大きくされており、容器体１への栓部材２の装着時には、第２筒状部２１は、径方向内方に圧縮されつつ第１筒状部１１に内嵌される。そして環状突起２１１が第１筒状部１１の内周面に密着して密閉する。なお、第２筒状部２１は、環状突起２１１を有さない構成としてもよい。環状突起２１１を有さない場合、第２筒状部２１の外周面が第１筒状部１１の内周面に面的に当接して密閉する。 The plug member 2 is for closing the opening 110 of the container body 1 and sealing the container body 1 . The plug member 2 is, for example, a molded rubber product, and includes a second tubular portion 21, a first flange portion 22, a first bottom portion 23 and a plurality of ports 24, as shown in FIGS. consists of The second cylindrical portion 21 has a substantially cylindrical shape, and contacts and seals the inner peripheral surface of the first cylindrical portion 11 . In this embodiment, a pair of annular protrusions 211 are formed on the outer peripheral portion of the second tubular portion 21 at intervals in the vertical direction (first direction). For example, the outer diameter dimension of the annular protrusion 211 in the natural state is slightly larger than the inner diameter dimension of the first cylindrical portion 11, and when the plug member 2 is attached to the container body 1, the second cylindrical portion 21 is It is fitted into the first cylindrical portion 11 while being compressed radially inward. Then, the annular protrusion 211 is in close contact with the inner peripheral surface of the first cylindrical portion 11 to seal it. Note that the second tubular portion 21 may be configured without the annular projection 211 . If the annular projection 211 is not provided, the outer peripheral surface of the second tubular portion 21 surface-contacts with the inner peripheral surface of the first tubular portion 11 for sealing.
　第１フランジ部２２は、概略円環状とされており、第２筒状部２１の上端（第１方向における一方側端）から径方向外方に延出している。第１フランジ部２２は、当該第１フランジ部２２の厚さ方向に見て第１筒状部１１と重なっており、第１筒状部１１の上端を塞いでいる。第１フランジ部２２は、適度な厚さを有し、上下方向（第１方向）の荷重に対して適度な弾性復元力を有する。第１フランジ部２２の厚さは、例えば１～３ｍｍ程度である。 The first flange portion 22 has a substantially annular shape and extends radially outward from the upper end (one side end in the first direction) of the second cylindrical portion 21 . The first flange portion 22 overlaps with the first tubular portion 11 when viewed in the thickness direction of the first flange portion 22 and closes the upper end of the first tubular portion 11 . The first flange portion 22 has an appropriate thickness and has an appropriate elastic restoring force against a load in the vertical direction (first direction). The thickness of the first flange portion 22 is, for example, about 1 to 3 mm.
　第１底部２３は、第２筒状部２１の下端（第１方向の他方側端）から径方向内方に延出しており、上下方向に見て第２筒状部２１の内側を塞いでいる。第１底部２３は、板状とされており、天井面２３１を有する。本実施形態において、天井面２３１は、下方（第１方向の他方側）を向く平坦面である。第１底部２３には、天井凹部２３２が形成されている。天井凹部２３２は、天井面２３１から上方（第１方向の一方側）に凹んだ部位である。天井凹部２３２は、第１底部２３の外周部寄りに設けられている。詳細は後述するが、天井凹部２３２は、ポート２４に対応して配置される。 The first bottom portion 23 extends radially inward from the lower end of the second tubular portion 21 (the other end in the first direction), and closes the inside of the second tubular portion 21 when viewed in the vertical direction. there is The first bottom portion 23 is plate-shaped and has a ceiling surface 231 . In this embodiment, the ceiling surface 231 is a flat surface facing downward (the other side in the first direction). A ceiling concave portion 232 is formed in the first bottom portion 23 . The ceiling recess 232 is a portion recessed upward (one side in the first direction) from the ceiling surface 231 . The ceiling recessed portion 232 is provided near the outer periphery of the first bottom portion 23 . Although the details will be described later, the ceiling recess 232 is arranged corresponding to the port 24 .
　本実施形態において、第１底部２３は、薄肉部２３８を有する。図４、図５等に表れているように、薄肉部２３８は、他の部位に比べて厚さが薄くされた部分である。薄肉部２３８は、第１底部２３の中央に配置されており、上下方向（第１方向）に見て略円形をなしている。本実施形態では、第１底部２３の中央部において上面が凹んだ形成とされており、当該第１底部２３の上面が凹んだ部位が薄肉部２３８とされる。図６において、薄肉部２３８と、厚さ方向（第１方向）に見て薄肉部２３８を囲む部位（以下、適宜「外周側厚肉部２３５」という）との境界を仮想線で表す。 In this embodiment, the first bottom portion 23 has a thin portion 238 . As shown in FIGS. 4, 5, etc., the thin portion 238 is a portion that is thinner than other portions. The thin portion 238 is arranged in the center of the first bottom portion 23 and has a substantially circular shape when viewed in the vertical direction (first direction). In the present embodiment, the top surface of the first bottom portion 23 is recessed at the central portion thereof, and the recessed portion of the top surface of the first bottom portion 23 is the thin portion 238 . In FIG. 6, the boundary between the thin portion 238 and a portion surrounding the thin portion 238 when viewed in the thickness direction (first direction) (hereinafter referred to as "peripheral thick portion 235") is indicated by a virtual line.
　本実施形態において、この薄肉部２３８は、ガス透過性を有する。本実施形態においてまた、薄肉部２３８は、透明である。薄肉部２３８の厚さは、例えば０．２～０．３ｍｍ程度である。薄肉部２３８を囲む外周側厚肉部２３５の厚さは、第１フランジ部２２の厚さと同程度とされており、例えば１～３ｍｍ程度である。 In this embodiment, the thin portion 238 has gas permeability. Also in this embodiment, thinned portion 238 is transparent. The thickness of the thin portion 238 is, for example, approximately 0.2 to 0.3 mm. The thickness of the outer peripheral thick portion 235 surrounding the thin portion 238 is about the same as the thickness of the first flange portion 22, for example, about 1 to 3 mm.
 複数のポート２４は、第１底部２３に設けられており、当該第１底部２３の適所に一体的に形成されている。本実施形態では、第１底部２３には一対のポート２４が設けられている。当該一対のポート２４は、第１底部２３の外周部寄り設けられており、上下方向（第１方向）に見て第１底部２３の中心を挟んで互いに反対側に配置されている。各ポート２４は、外周側厚肉部２３５に設けられている。 A plurality of ports 24 are provided on the first bottom portion 23 and are integrally formed at appropriate locations on the first bottom portion 23 . In this embodiment, the first bottom portion 23 is provided with a pair of ports 24 . The pair of ports 24 are provided near the outer periphery of the first bottom portion 23 and are arranged on opposite sides of the center of the first bottom portion 23 when viewed in the vertical direction (first direction). Each port 24 is provided in the outer thick portion 235 .
　ポート２４は、容器体１の内部に培養液を供給し、あるいは容器体１内の培養液を外部に排出するためのものであり、図４に示すように延出部２４１および流路２４２を有する。延出部２４１は、第１底部２３から上方（第１方向の一方側）に延びる円筒状とされており、横断面円形の外周面２４１ａを有する。本実施形態において、延出部２４１の先端には、外周面２４１ａよりも外向きに突出する突起部２４５が設けられている。 The port 24 is for supplying the culture solution to the inside of the container body 1 or discharging the culture solution inside the container body 1 to the outside, and as shown in FIG. have. The extending portion 241 has a cylindrical shape extending upward (one side in the first direction) from the first bottom portion 23 and has an outer peripheral surface 241a having a circular cross section. In the present embodiment, a projection 245 is provided at the tip of the extension 241 and projects outward from the outer peripheral surface 241a.
　流路２４２は、上端（第１方向の一方側端）が外部に開放し、かつ下端（第１方向の手方側端）が容器体１の内側空間において開放しており、外部と容器体１の内側空間とに通じる連通路である。本実施形態では、流路２４２は、横断面円形とされており、延出部２４１に形成される。 The flow path 242 has an upper end (one side end in the first direction) open to the outside and a lower end (hand side end in the first direction) open to the inner space of the container body 1 so that the outside and the container body are open. It is a communication passage leading to the inner space of 1. In this embodiment, the channel 242 has a circular cross section and is formed in the extension 241 .
　天井凹部２３２は、上下方向（第１方向）に見てポート２４と重なり、流路２４２につながっている。本実施形態では、天井凹部２３２は、錐形状とされており、流路２４２とつながる位置から下方に向かうにつれて横断面の面積が大きくなるように形成されている。本実施形態で図示した例では、天井凹部２３２は、第１底部２３からポート２４（延出部２４１）に跨って形成されている。 The ceiling recessed portion 232 overlaps the port 24 when viewed in the vertical direction (first direction) and is connected to the flow path 242 . In this embodiment, the ceiling recessed portion 232 has a conical shape, and is formed such that the area of the cross section increases downward from the position where it is connected to the flow path 242 . In the illustrated example of the present embodiment, the ceiling recess 232 is formed across the first bottom portion 23 and the port 24 (extending portion 241).
　上記構成の栓部材２は、可撓性、弾力性を有する軟質素材からなる。栓部材２を構成する素材としては、例えば、シリコーンゴム、エラストマー樹脂などが挙げられる。詳細は後述するが、栓部材２と内容物（例えば培養液）との接触を考慮すると、栓部材２の素材としては、細胞毒性が無く、かつ生体適合性を有する医療用シリコーンゴムがより好ましい。また、栓部材２の硬さについては、例えばゴム硬度が２０度～４０度程度であるのが好ましい。 The plug member 2 configured as described above is made of a soft material having flexibility and elasticity. Examples of the material forming the plug member 2 include silicone rubber and elastomer resin. Although the details will be described later, in consideration of the contact between the plug member 2 and the contents (for example, culture solution), the material of the plug member 2 is more preferably non-cytotoxic and biocompatible medical silicone rubber. . As for the hardness of the plug member 2, it is preferable that the rubber hardness is about 20 to 40 degrees, for example.
　カバー部材３は、栓部材２に重なって当該栓部材２に当接するものである。本実施形態において、カバー部材３は、第２フランジ部３１、第４筒状部３２、第２底部３３および第５筒状部３４を含んで構成される。 The cover member 3 overlaps the plug member 2 and contacts the plug member 2 . In this embodiment, the cover member 3 includes a second flange portion 31 , a fourth tubular portion 32 , a second bottom portion 33 and a fifth tubular portion 34 .
　図４、図５に示すように、第２フランジ部３１は、第１フランジ部２２を上方から覆っており、当該第１フランジ部２２の上面に当接している。第４筒状部３２は、第２フランジ部３１の内周縁につながっている。第４筒状部３２は、概略円筒状とされており、第２フランジ部３１の内周縁から下方（第１方向の他方側）に延びている。第４筒状部３２は、第２筒状部２１の内周面を覆っており、当該第２筒状部２１の内周面に当接している。 As shown in FIGS. 4 and 5, the second flange portion 31 covers the first flange portion 22 from above and is in contact with the upper surface of the first flange portion 22 . The fourth tubular portion 32 is connected to the inner peripheral edge of the second flange portion 31 . The fourth tubular portion 32 has a substantially cylindrical shape and extends downward (the other side in the first direction) from the inner peripheral edge of the second flange portion 31 . The fourth tubular portion 32 covers the inner peripheral surface of the second tubular portion 21 and is in contact with the inner peripheral surface of the second tubular portion 21 .
　第２底部３３は、第４筒状部３２の下端（第１方向の他方側端）につながり、当該第４筒状部３２の下端から径方向内方に延びている。第２底部３３は、第１底部２３を上方から覆っており、当該第１底部２３の上面に当接している。本実施形態では、第２底部３３の中央部は、薄肉部２３８の形成箇所に対応して下方（第１方向の他方側）に凹んだ形状とされている。 The second bottom portion 33 is connected to the lower end of the fourth tubular portion 32 (the other end in the first direction) and extends radially inward from the lower end of the fourth tubular portion 32 . The second bottom portion 33 covers the first bottom portion 23 from above and is in contact with the upper surface of the first bottom portion 23 . In this embodiment, the central portion of the second bottom portion 33 is recessed downward (on the other side in the first direction) corresponding to the location where the thin portion 238 is formed.
　第２底部３３には、貫通孔３３１が形成されている。貫通孔３３１は、第２底部３３を厚さ方向に貫通しており、上下方向（第１方向）に見て栓部材２の薄肉部２３８と重なっている。本実施形態では、第２底部３３には複数の貫通孔３３１が形成されている。図３等に示すように、複数の貫通孔３３１は、上下方向に見て適度に分散して配置されている。 A through hole 331 is formed in the second bottom portion 33 . The through hole 331 penetrates the second bottom portion 33 in the thickness direction and overlaps the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction). In this embodiment, a plurality of through holes 331 are formed in the second bottom portion 33 . As shown in FIG. 3 and the like, the plurality of through holes 331 are appropriately distributed when viewed in the vertical direction.
　第５筒状部３４は、第２底部３３に設けられており、当該第２底部３３の適所から上方（第１方向の一方側）に延びている。本実施形態において、第２底部３３には一対の第５筒状部３４が設けられている。当該一対の第５筒状部３４は、上記一対のポート２４それぞれに対応して配置されている。本実施形態では、各第５筒状部３４は、第４筒状部３２の一部につながる形状とされている。一対の第５筒状部３４それぞれには、ポート２４（延出部２４１）が嵌挿されている。具体的には、第５筒状部３４の内径寸法は、自然状態において延出部２４１の外径寸法よりも僅かに小さくされている。各延出部２４１は、第５筒状部３４に圧入されている。これにより、第５筒状部３４は、延出部２４１の外周面２４１ａに密着（当接）して当該外周面２４１ａを囲っている。 The fifth tubular portion 34 is provided on the second bottom portion 33 and extends upward (one side in the first direction) from a suitable location on the second bottom portion 33 . In this embodiment, the second bottom portion 33 is provided with a pair of fifth cylindrical portions 34 . The pair of fifth cylindrical portions 34 are arranged corresponding to the pair of ports 24, respectively. In this embodiment, each fifth tubular portion 34 is shaped to be connected to a portion of the fourth tubular portion 32 . The port 24 (extending portion 241) is fitted in each of the pair of fifth cylindrical portions 34. As shown in FIG. Specifically, the inner diameter dimension of the fifth tubular portion 34 is slightly smaller than the outer diameter dimension of the extending portion 241 in the natural state. Each extending portion 241 is press-fitted into the fifth tubular portion 34 . As a result, the fifth tubular portion 34 is in close contact with (contacts with) the outer peripheral surface 241a of the extending portion 241 to surround the outer peripheral surface 241a.
　また、本実施形態では、図４等から理解されるように、上下方向（第１方向）に見て、第５筒状部３４は、ポート２４の先端に設けられた突起部２４５と重なっている。突起部２４５は、第５筒状部３４の上方に隣接している。これにより、第５筒状部３４の上端には、突起部２４５が当接し得る。 In addition, in the present embodiment, as can be understood from FIG. 4 and the like, when viewed in the vertical direction (first direction), the fifth cylindrical portion 34 overlaps the protrusion 245 provided at the tip of the port 24. there is The projecting portion 245 is adjacent above the fifth tubular portion 34 . As a result, the protrusion 245 can come into contact with the upper end of the fifth tubular portion 34 .
　カバー部材３は、硬質素材からなり、例えば半透明または透明のプラスチック材料により形成されている。当該プラスチック材料としては、例えばポリスチレンやメチルペンテンのほか、ポリカーボネート、シクロオレフィンポリマー、シクロオレフィンコポリマーなどの、好適には透明性を有する材料が用いられるが、これらに限定されない。 The cover member 3 is made of a hard material, such as a translucent or transparent plastic material. Examples of the plastic material include, but are not limited to, polystyrene, methylpentene, polycarbonate, cycloolefin polymer, cycloolefin copolymer, and other transparent materials.
　外蓋４は、頂板部４１および第３筒状部４２を有する。頂板部４１は、概略円環状とされており、カバー部材３の第２フランジ部３１を容器体１の第１筒状部１１の上端１１４との間に挟み付ける部分である。頂板部４１の径方向内側には、貫通孔４１１が形成されている。貫通孔４１１は、頂板部４１の厚さ方向視において当該頂板部４１に囲まれている。頂板部４１の厚さ方向（上下方向）に見て、貫通孔４１１は、栓部材２の薄肉部２３８およびカバー部材３における複数の貫通孔３３１と重なっている。 The outer lid 4 has a top plate portion 41 and a third tubular portion 42 . The top plate portion 41 has a substantially annular shape and is a portion that sandwiches the second flange portion 31 of the cover member 3 and the upper end 114 of the first cylindrical portion 11 of the container body 1 . A through hole 411 is formed radially inside the top plate portion 41 . The through hole 411 is surrounded by the top plate portion 41 when viewed in the thickness direction of the top plate portion 41 . The through hole 411 overlaps the thin portion 238 of the plug member 2 and the plurality of through holes 331 of the cover member 3 when viewed in the thickness direction (vertical direction) of the top plate portion 41 .
　第３筒状部４２は、頂板部４１の外周縁から当該頂板部４１の厚さ方向（図中下方）に延びており、概略円筒状とされている。第３筒状部４２の内周面には、雌ネジ４２１が形成されており、当該雌ネジ４２１は、容器体１（第１筒状部１１）の雄ネジ１１１に螺合可能である。 The third tubular portion 42 extends from the outer peripheral edge of the top plate portion 41 in the thickness direction (downward in the figure) of the top plate portion 41 and has a substantially cylindrical shape. A female screw 421 is formed on the inner peripheral surface of the third tubular portion 42, and the female screw 421 can be screwed into the male screw 111 of the container body 1 (first tubular portion 11).
　本実施形態において、頂板部４１には、凸状部４１２が設けられている。図５によく表れているように、凸状部４１２は、頂板部４１の厚さ方向を向く主面４１ａから上方（第１方向の一方側）に突き出ている。本実施形態において凸状部４１２は、互いに分離して複数設けられている。これら凸状部４１２の先端４１２ａがなす平面Ｐ１は、頂板部４１の主面４１ａと略平行である。ここで、「略平行」とは、平面Ｐ１と主面４１ａとが平行となるように設計されるが、寸法公差等により多少の平行度のバラつきがある場合も含むものである。また、複数に凸状部４１２の各先端４１２ａがなす平面Ｐ１は、細胞培養容器Ａ１において最も上方に位置する。図１～図５から理解されるように、本実施形態において、凸状部４１２は、上下方向視において同心円の円弧状に設けられ、隣接する凸状部４１２の間が空隙となっている。これにより、凸状部４１２は、上下方向（頂板部４１の厚さ方向）に見て閉じた環状を形成しない。 In this embodiment, the top plate portion 41 is provided with a convex portion 412 . As shown in FIG. 5, the convex portion 412 protrudes upward (one side in the first direction) from the main surface 41a of the top plate portion 41 facing the thickness direction. In this embodiment, a plurality of convex portions 412 are provided separately from each other. A plane P<b>1 formed by the tips 412 a of these convex portions 412 is substantially parallel to the main surface 41 a of the top plate portion 41 . Here, "substantially parallel" is designed so that the plane P1 and the main surface 41a are parallel, but it also includes the case where there is some variation in parallelism due to dimensional tolerance or the like. A plane P1 formed by the tips 412a of the plurality of convex portions 412 is located at the highest position in the cell culture container A1. As can be understood from FIGS. 1 to 5, in the present embodiment, the convex portions 412 are provided in the shape of concentric arcs when viewed in the up-down direction, and gaps are formed between adjacent convex portions 412 . As a result, the convex portion 412 does not form a closed annular shape when viewed in the vertical direction (thickness direction of the top plate portion 41).
　なお、図４、図５に表れているように、容器体１の容器底部１２には、その外周縁から下方に突出するスカート部１２１がつながっている。そして、複数の凸状部４１２により形成される同心円円弧の外周縁の外径寸法は、スカート部１２１の内径寸法よりも僅かに小さくされている。このような構成によれば、細胞培養容器Ａ１の凸状部４１２が、他の細胞培養容器Ａ１のスカート部１２１に嵌まるようにして当該他の細胞培養容器Ａ１を重ねることができる。したがって、複数の細胞培養容器Ａ１を安定姿勢で積み重ねることができる。 As shown in FIGS. 4 and 5, the container bottom portion 12 of the container body 1 is connected to a skirt portion 121 projecting downward from the outer peripheral edge thereof. The outer diameter dimension of the outer peripheral edge of the concentric arcs formed by the plurality of convex portions 412 is slightly smaller than the inner diameter dimension of the skirt portion 121 . According to such a configuration, the convex portion 412 of the cell culture vessel A1 can be fitted into the skirt portion 121 of the other cell culture vessel A1 so that the other cell culture vessel A1 can be stacked. Therefore, a plurality of cell culture vessels A1 can be stacked in a stable posture.
　外蓋４は、例えば不透明、半透明または透明のプラスチック材料により形成されている。当該プラスチック材料としては、例えばポリエチレン、ポリプロピレンのほか、ポリスチレンやメチルペンテン、ポリカーボネートなどが用いられるが、これらに限定されない。 The outer lid 4 is made of, for example, an opaque, translucent or transparent plastic material. Examples of the plastic material include, but are not limited to, polyethylene, polypropylene, polystyrene, methylpentene, and polycarbonate.
　次に、細胞培養容器Ａ１の使用方法および作用について、図７～図９を参照しつつ説明する。 Next, the usage method and action of the cell culture vessel A1 will be described with reference to FIGS. 7 to 9.
　細胞培養容器Ａ１は、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。なお、細胞培養容器Ａ１（容器体１）に収容される培養細胞や培養液については、特に限定されるものではない。 The cell culture vessel A1 is used for perfusion culture in which the culture solution is automatically supplied at a constant low speed and the same amount of culture solution is discharged at the same time. It should be noted that the cultured cells and the culture solution accommodated in the cell culture container A1 (container body 1) are not particularly limited.
　細胞培養容器Ａ１を用いた細胞培養では、あらかじめ栓部材２とカバー部材３、各ポート２４とコネクタ７をアセンブリしておく(図７参照)。コネクタ７は、一対のポート２４それぞれに圧入接続される。一方のポート２４には、コネクタ７およびチューブ８を介して灌流用ポンプ（図示略）が接続され、他方のポート２４には、コネクタ７およびチューブ８を介して排液用容器（図示略）が接続される。なお、各チューブ８は、あらかじめ外蓋４の内側を通しておく。次に、容器体１内に培養する細胞を播種し、容器体１内に培養液を所定量入れる。次いで、容器体１に栓部材２とカバー部材３を被せ、外蓋４で締め付ける。 In the cell culture using the cell culture vessel A1, the stopper member 2, the cover member 3, each port 24 and the connector 7 are assembled in advance (see Fig. 7). The connector 7 is press-fitted to each of the pair of ports 24 . A perfusion pump (not shown) is connected to one port 24 via a connector 7 and a tube 8, and a drainage container (not shown) is connected to the other port 24 via a connector 7 and a tube 8. Connected. Each tube 8 is passed through the inner side of the outer lid 4 in advance. Next, cells to be cultured are seeded in the container body 1, and a predetermined amount of culture solution is placed in the container body 1. FIG. Next, the container body 1 is covered with the plug member 2 and the cover member 3, and the outer lid 4 is tightened.
　続いて、一対のポート２４およびこれらに接続されたコネクタ７を介して、細胞培養容器Ａ１に対して培養液の供給および空気の排出を行い、容器体１内を培養液で満たす。ここで、一方のポート２４を通じて培養液を容器体１内に供給しつつ、他方のポート２４を通じて容器体１内の空気を外部に排出する。図８は、容器体１内に培養液Ｃを満たした状態を示す。培養液Ｃは、容器体１の内側空間（容器体１の容器底部１２および第１筒状部１１と、栓部材２の第１底部２３とで囲まれた空間）を満たす。このとき、容器体１の上記内側空間に少量の空気（以下、適宜「気泡Ｂ」という）が残存する場合がある。 Subsequently, via the pair of ports 24 and the connector 7 connected thereto, the culture medium is supplied to the cell culture vessel A1 and the air is discharged, thereby filling the container body 1 with the culture medium. Here, the culture solution is supplied into the vessel 1 through one port 24 and the air inside the vessel 1 is discharged to the outside through the other port 24 . FIG. 8 shows a state in which the container body 1 is filled with the culture medium C. As shown in FIG. The culture solution C fills the inner space of the container body 1 (the space surrounded by the container bottom portion 12 and the first cylindrical portion 11 of the container body 1 and the first bottom portion 23 of the plug member 2). At this time, a small amount of air (hereinafter referred to as “bubble B” as appropriate) may remain in the inner space of the container body 1 .
　細胞培養容器Ａ１において、栓部材２の第１底部２３における天井面２３１は平坦面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２はポート２４の流路２４２につながっている。このような構成によれば、図９に示すように、ポート２４が上位に位置するように細胞培養容器Ａ１を傾けると、容器体１内の気泡Ｂは、天井面２３１を伝ってポート２４付近に移動する。この状態で灌流用ポンプを作動させることにより、容器体１内に残存する気泡Ｂはさらに天井凹部２３２を通じて流路２４２内に移動し、細胞培養容器Ａ１の外部に排出することができる。そして、灌流培養を行う際には、一方のポート２４を通じて新しい培養液Ｃが容器体１（細胞培養容器Ａ１）に供給され、かつ他方のポート２４を通じて容器体１内（細胞培養容器Ａ１内部）の培養液Ｃが外部に排出される。灌流培養工程においては、容器体１内の培養液Ｃを僅かずつ入れ替え続ける。 In the cell culture container A1, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is a flat surface. A ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, as shown in FIG. 9, when the cell culture container A1 is tilted so that the port 24 is positioned at the top, the air bubbles B in the container body 1 travel along the ceiling surface 231 and near the port 24. move to By operating the perfusion pump in this state, the air bubbles B remaining in the container body 1 can be further moved into the channel 242 through the ceiling recess 232 and discharged to the outside of the cell culture container A1. When performing perfusion culture, a new culture solution C is supplied to the container body 1 (cell culture container A1) through one port 24, and the container body 1 (inside the cell culture container A1) is supplied through the other port 24. of the culture solution C is discharged to the outside. In the perfusion culture process, the culture solution C in the vessel body 1 is continuously replaced little by little.
　細胞培養容器Ａ１は、容器体１および栓部材２の他に、外蓋４を備えている。容器体１の第１筒状部１１に外嵌される外蓋４の第３筒状部４２には、雌ネジ４２１が形成されている。当該雌ネジ４２１を容器体１（第１筒状部１１）の雄ネジ１１１に締め込むことで、栓部材２における第１フランジ部２２が、第１筒状部１１の上端１１４と外蓋４の頂板部４１とによって挟まれて、アセンブリ状態が維持される。栓部材２は、外周側の第１フランジ部２２から中央の第１底部２３ないし薄肉部２３８に至る一連の膜状である。これにより、容器体１に収容された内容物（培養細胞や培養液Ｃ）は液封されている。したがって、灌流培養工程において細胞培養容器Ａ１の閉鎖状態を維持したまま培養液灌流作業を行うことができ、コンタミネーションのリスクを回避することができる。したがって、細胞培養容器Ａ１によれば、培養液Ｃの状態を良好に保って、細胞の品質確保を実現することができる。なお、容器体１（第１筒状部１１）の雄ネジ１１１および外蓋４（第３筒状部４２）の雌ネジ２２１は、本開示の「係止手段」の一例に相当する。 The cell culture vessel A1 has an outer lid 4 in addition to the vessel body 1 and the plug member 2. A female screw 421 is formed in the third tubular portion 42 of the outer lid 4 fitted onto the first tubular portion 11 of the container body 1 . By tightening the female thread 421 into the male thread 111 of the container body 1 (first cylindrical portion 11 ), the first flange portion 22 of the plug member 2 is connected to the upper end 114 of the first cylindrical portion 11 and the outer lid 4 . The assembled state is maintained by being sandwiched between the top plate portion 41 of the . The plug member 2 is in the form of a series of films extending from the first flange portion 22 on the outer peripheral side to the central first bottom portion 23 to the thin portion 238 . As a result, the contents (cultured cells and culture medium C) housed in the container body 1 are liquid-sealed. Therefore, in the perfusion culture process, the culture solution perfusion work can be performed while maintaining the closed state of the cell culture vessel A1, and the risk of contamination can be avoided. Therefore, according to the cell culture vessel A1, it is possible to keep the culture solution C in good condition and ensure the quality of the cells. The male screw 111 of the container body 1 (first tubular portion 11) and the female screw 221 of the outer lid 4 (third tubular portion 42) correspond to an example of the "locking means" of the present disclosure.
　細胞培養容器Ａ１は、容器体１および栓部材２の他に、カバー部材３を備えている。栓部材２は、軟質素材からなる。カバー部材３は、硬質素材からなり、栓部材２に重なって当接している。具体的には、カバー部材３は、栓部材２の第１フランジ部２２を覆う第２フランジ部３１を含む。このような構成によれば、第２フランジ部３１は栓部材２の第１フランジ部２２と外蓋４の頂板部４１との間に介在する。したがって、外蓋４の雌ネジ４２１を容器体１（第１筒状部１１）の雄ネジ１１１に締め込む際、頂板部４１が第２フランジ部３１（硬質素材）と摺接し、外蓋４の摺動性が良好となる。 The cell culture vessel A1 includes a cover member 3 in addition to the container body 1 and the plug member 2. The plug member 2 is made of a soft material. The cover member 3 is made of a hard material and is in contact with the plug member 2 in an overlapping manner. Specifically, the cover member 3 includes a second flange portion 31 that covers the first flange portion 22 of the plug member 2 . With such a configuration, the second flange portion 31 is interposed between the first flange portion 22 of the plug member 2 and the top plate portion 41 of the outer lid 4 . Therefore, when the female screw 421 of the outer lid 4 is screwed into the male screw 111 of the container body 1 (first cylindrical portion 11), the top plate portion 41 is in sliding contact with the second flange portion 31 (hard material). slidability becomes good.
　カバー部材３は、第２筒状部２１の内周面を覆う第４筒状部３２を含む。このような構成によれば、容器体１（第１筒状部１１）の内周面に軟質素材からなる栓部材２が押し付けられ、細胞培養容器Ａ１のシール性が高められる。また、カバー部材３は、栓部材２の第１底部２３を覆う第２底部３３を含む。このような構成によれば、灌流培養工程において、第１底部２３（薄肉部２３８）は、供給される培養液Ｃの加圧力によって上方に膨らもうとするが、第２底部３３によって第１底部２３（薄肉部２３８）の変形が抑制される。これにより、容器体１内の培養液Ｃの流れがスムーズになるとともに、容器体１内部の視認性にも優れ、容器体１内の状況を観察するのに適する。 The cover member 3 includes a fourth tubular portion 32 that covers the inner peripheral surface of the second tubular portion 21 . According to such a configuration, the plug member 2 made of a soft material is pressed against the inner peripheral surface of the container body 1 (first tubular portion 11), and the sealing performance of the cell culture container A1 is enhanced. The cover member 3 also includes a second bottom portion 33 that covers the first bottom portion 23 of the plug member 2 . According to such a configuration, in the perfusion culture process, the first bottom portion 23 (thin portion 238) tries to swell upward due to the pressurizing force of the supplied culture medium C, but the second bottom portion 33 causes the first bottom portion 23 to expand. Deformation of the bottom portion 23 (thin portion 238) is suppressed. As a result, the flow of the culture solution C inside the container body 1 becomes smooth, and the visibility inside the container body 1 is excellent, making it suitable for observing the conditions inside the container body 1 .
　栓部材２に設けられたポート２４は、第１底部２３から上方に延びる延出部２４１を有する。また、カバー部材３は第５筒状部３４を含み、第５筒状部３４は、延出部２４１の外周面２４１ａに当接して当該外周面２４１ａを覆う。このような構成によれば、ポート２４（延出部２４１）にコネクタ７が圧入接続される場合、延出部２４１が径方向外方に膨らむことは、第５筒状部３４によって抑制される。その結果、ポート２４へのコネクタ７接続時のシール性が高められる。 The port 24 provided on the plug member 2 has an extending portion 241 extending upward from the first bottom portion 23 . Further, the cover member 3 includes a fifth cylindrical portion 34, and the fifth cylindrical portion 34 contacts the outer peripheral surface 241a of the extending portion 241 to cover the outer peripheral surface 241a. According to such a configuration, when the connector 7 is press-fitted into the port 24 (the extension portion 241), the extension portion 241 is prevented from expanding radially outward by the fifth tubular portion 34. . As a result, the sealing performance when connecting the connector 7 to the port 24 is enhanced.
　栓部材２の第１底部２３は、ガス透過性を有する薄肉部２３８を含む。このため、容器体１（細胞培養容器Ａ１）の内容物について、当該容器体１（細胞培養容器Ａ１）の外部との通気状態が維持される。したがって、本実施形態によれば、容器体１（細胞培養容器Ａ１）の内容物について、外部と通気状態での培養を行うことができる。また、細胞培養容器Ａ１を所定のガス雰囲気下にあるインキュベータ内に配置すれば、当該インキュベータ内のガス雰囲気を容器体１（細胞培養容器Ａ１）の内部に取り込むことが可能である。 The first bottom portion 23 of the plug member 2 includes a thin portion 238 having gas permeability. Therefore, the contents of the container body 1 (cell culture container A1) are maintained in a ventilated state with the outside of the container body 1 (cell culture container A1). Therefore, according to the present embodiment, the contents of the container body 1 (cell culture container A1) can be cultured in an aerated state with the outside. Further, if the cell culture vessel A1 is placed in an incubator under a predetermined gas atmosphere, the gas atmosphere in the incubator can be taken into the container body 1 (cell culture vessel A1).
　カバー部材３の第２底部３３には、当該第２底部３３の厚さ方向に貫通する貫通孔３３１が形成されている。貫通孔３３１は、上下方向（第１方向）に見て栓部材２の薄肉部２３８と重なっている。このような構成によれば、上記のように第１底部２３（薄肉部２３８）の変形を抑制しつつ、薄肉部２３８を介して容器体１内と外部との通気状態を維持することができる。 A through hole 331 is formed in the second bottom portion 33 of the cover member 3 so as to pass through the second bottom portion 33 in the thickness direction. The through hole 331 overlaps the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction). According to such a configuration, while suppressing the deformation of the first bottom portion 23 (the thin portion 238 ) as described above, it is possible to maintain the state of ventilation between the inside and the outside of the container body 1 via the thin portion 238 . .
　ポート２４（延出部２４１）の先端には、延出部２４１の外周面２４１ａよりも外向きに突出する突起部２４５が設けられている。突起部２４５は、第５筒状部３４の上方に隣接しており、第５筒状部３４の上端に当接し得る。このような構成によれば、延出部２４１に下方への押付力が作用しても、延出部２４１が第５筒状部３４を通じて容器体１の内側空間に押し込まれることは防止される。これにより、例えばコネクタ７をポート２４に圧入接続する際、ポート２４に下方への押圧力が作用するが、容器体１の内側空間への延出部２４１の不当な侵入が阻止され、容器体１内の細胞の損傷等が回避される。 A protrusion 245 that protrudes outward from the outer peripheral surface 241 a of the extension 241 is provided at the tip of the port 24 (extension 241 ). The projecting portion 245 is adjacent to the upper side of the fifth tubular portion 34 and can abut on the upper end of the fifth tubular portion 34 . According to such a configuration, even if a downward pressing force acts on the extending portion 241, the extending portion 241 is prevented from being pushed into the inner space of the container body 1 through the fifth cylindrical portion 34. . As a result, when the connector 7 is press-fitted to the port 24, for example, a downward pressing force acts on the port 24, but the extending portion 241 is prevented from entering the inner space of the container body 1 unreasonably. Damage to cells in 1 is avoided.
＜第１実施形態の第１変形例＞
　図１０は、第１実施形態の第１変形例に係る細胞培養容器を示している。なお、図１０以降の図面において、上記実施形態の細胞培養容器Ａ１と同一または類似の要素には、上記実施形態と同一の符号を付しており、適宜説明を省略する。 <First Modification of First Embodiment>
FIG. 10 shows a cell culture vessel according to a first modified example of the first embodiment. In addition, in the drawings after FIG. 10 , the same or similar elements as those of the cell culture vessel A1 of the above embodiment are denoted by the same reference numerals as those of the above embodiment, and description thereof will be omitted as appropriate.
　本変形例の細胞培養容器Ａ１１においては、主に天井凹部２３２の構成が上記実施形態の細胞培養容器Ａ１と異なっている。 The cell culture vessel A11 of this modified example differs from the cell culture vessel A1 of the above embodiment mainly in the configuration of the ceiling recess 232 .
　本変形例において、天井凹部２３２は、錐形状部２３３および周縁部２３４を有する。錐形状部２３３は、流路２４２につながっている。図示した例では、錐形状部２３３は、流路２４２とつながる位置から下方に向かうにつれて横断面の面積が大きくなるように形成された円錐形状部分である。周縁部２３４は、錐形状部２３３につながり、かつ第１底部２３の外周縁２３７に通じている。周縁部２３４は、第１底部２３の径方向外方に向かうにつれて下方に位置するように傾斜している。図１１に表れているように、周縁部２３４は、第１底部２３の厚さ方向（第１方向）に見て、径方向外方に拡がる概略扇形である。また、本変形例では、第２筒状部２１の外周部には環状突起２１１が形成されておらず、第２筒状部２１の外周面が第１筒状部１１の内周面に面的に当接している。 In this modified example, the ceiling concave portion 232 has a conical portion 233 and a peripheral edge portion 234 . The conical portion 233 is connected to the channel 242 . In the illustrated example, the cone-shaped portion 233 is a cone-shaped portion formed so that the cross-sectional area increases downward from the position where it connects with the flow path 242 . The peripheral edge portion 234 connects to the conical portion 233 and communicates with the outer peripheral edge 237 of the first bottom portion 23 . The peripheral edge portion 234 is inclined downward as it goes radially outward of the first bottom portion 23 . As shown in FIG. 11 , the peripheral edge portion 234 has a substantially fan shape extending radially outward when viewed in the thickness direction (first direction) of the first bottom portion 23 . Further, in this modification, the annular protrusion 211 is not formed on the outer peripheral portion of the second tubular portion 21 , and the outer peripheral surface of the second tubular portion 21 faces the inner peripheral surface of the first tubular portion 11 . are in direct contact.
　本変形例の細胞培養容器Ａ１１は、培養細胞と培養液を収容し、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。細胞培養容器Ａ１１においては、栓部材２の第１底部２３における天井面２３１は平坦面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２はポート２４の流路２４２につながっている。このような構成によれば、灌流培養に先立ち容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、ポート２４が上位に位置するように細胞培養容器Ａ１１を傾けると、容器体１内の気泡は、天井面２３１を伝ってポート２４付近に移動する。そして、当該気泡は、天井凹部２３２を通じて流路２４２内を浮上する。これにより、容器体１内に残存する気泡を細胞培養容器Ａ１１の外部に排出することができる。 The cell culture vessel A11 of this modified example is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time. In the cell culture vessel A11, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. A ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position. When the cell culture container A11 is tilted, air bubbles in the container body 1 move along the ceiling surface 231 to the vicinity of the port 24 . Then, the air bubbles float in the channel 242 through the ceiling recess 232 . As a result, air bubbles remaining in the vessel body 1 can be discharged to the outside of the cell culture vessel A11.
　本変形例では、天井凹部２３２（周縁部２３４）は、第１底部２３の外周縁２３７に通じている。このような構成によれば、容器体１の内側空間に残存する気泡を排出するために細胞培養容器Ａ１１を傾けた際、気泡が第１底部２３の外周縁２３７側に到達しても、当該気泡を、周縁部２３４を伝わせて流路２４２に導くことができる。これにより、容器体１の内側空間に残存する気泡を、より的確に排出することが可能である。 In this modified example, the ceiling recessed portion 232 (peripheral edge portion 234 ) communicates with the outer peripheral edge 237 of the first bottom portion 23 . According to such a configuration, when the cell culture container A11 is tilted in order to discharge the air bubbles remaining in the inner space of the container body 1, even if the air bubbles reach the outer peripheral edge 237 side of the first bottom portion 23, Air bubbles can be directed to channel 242 along peripheral edge 234 . Thereby, it is possible to more accurately discharge the air bubbles remaining in the inner space of the container body 1 .
　その他にも、本変形例の細胞培養容器Ａ１１において、上記の細胞培養容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A11 of this modified example has the same effects as the cell culture vessel A1 described above.
　図１２は、栓部材２の他の例を示す底面図である。図１２に示した栓部材２においては、図１１に示した栓部材２と比べて、周縁部２３４の形状が異なる。図１２に示した例では、周縁部２３４は、第１底部２３の外周部において円周方向に沿って延びるように形成されている。図１２に示した周縁部２３４は、図１１に示した周縁部２３４と同様に、錐形状部２３３につながり、かつ第１底部２３の外周縁２３７に通じている。また、周縁部２３４は、第１底部２３の径方向外方に向かうにつれて下方に位置するように傾斜している。 12 is a bottom view showing another example of the plug member 2. FIG. The plug member 2 shown in FIG. 12 differs from the plug member 2 shown in FIG. 11 in the shape of the peripheral portion 234 . In the example shown in FIG. 12 , the peripheral edge portion 234 is formed to extend along the circumferential direction on the outer peripheral portion of the first bottom portion 23 . The peripheral edge 234 shown in FIG. 12, like the peripheral edge 234 shown in FIG. In addition, the peripheral edge portion 234 is inclined so as to be positioned downward as it goes radially outward of the first bottom portion 23 .
　詳細な図示説明は省略するが、図１２に示した栓部材２を備えて構成された細胞培養容器においても、上記の細胞培養容器Ａ１１と同様に、容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、当該気泡を、周縁部２３４を伝わせて流路２４２に導くことができる。これにより、容器体１の内側空間に残存する気泡を、より的確に排出することが可能である。 Although detailed illustration and explanation are omitted, in the cell culture vessel configured with the plug member 2 shown in FIG. ), even if air bubbles remain in the inner space, the air bubbles can be guided to the flow path 242 along the peripheral edge portion 234 . Thereby, it is possible to more accurately discharge the air bubbles remaining in the inner space of the container body 1 .
＜第１実施形態の第２変形例＞
　図１３は、第１実施形態の第２変形例に係る細胞培養容器を示している。本変形例の細胞培養容器Ａ１２においては、栓部材２の第１底部２３およびカバー部材３の第２底部３３の構成が上記実施形態の細胞培養容器Ａ１と異なっている。本変形例において、第１底部２３（主に薄肉部２３８）は、湾曲する板状とされており、径方向内方に向かうにつれて下方（第１方向の他方側）に位置するように湾曲している。天井面２３１は、湾曲面であり、径方向内方に向かうにつれて下方に位置するよう湾曲している。本変形例では、第２底部３３についても、径方向内方に向かうにつれて下方（第１方向の他方側）に位置するよう湾曲している。上記実施形態の細胞培養容器Ａ１において、第２底部３３は上下方向（第１方向）に見て栓部材２の薄肉部２３８と重なる部分が平板状とされていた（図４参照）。本変形例では、第２底部３３は、上下方向（第１方向）に見て栓部材２の薄肉部２３８と重なる部分が湾曲状とされている。第２底部３３の下面は、薄肉部２３８と当接している。なお、薄肉部２３８については、自然状態において湾曲形状であってもよいが、これに限らない。薄肉部２３８は、例えば自然状態において平板状とされ、カバー部材３の第２底部３３が被せられることで湾曲形状となるように構成してもよい。 <Second Modification of First Embodiment>
FIG. 13 shows a cell culture vessel according to a second modification of the first embodiment. In the cell culture vessel A12 of this modified example, the configurations of the first bottom portion 23 of the plug member 2 and the second bottom portion 33 of the cover member 3 are different from those of the cell culture vessel A1 of the above embodiment. In this modification, the first bottom portion 23 (mainly the thin-walled portion 238) has a curved plate shape, and curves downward (on the other side in the first direction) as it extends radially inward. ing. The ceiling surface 231 is a curved surface that curves downward as it extends radially inward. In this modified example, the second bottom portion 33 is also curved so as to be positioned downward (on the other side in the first direction) as it goes radially inward. In the cell culture vessel A1 of the above embodiment, the portion of the second bottom portion 33 that overlaps the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction) is flat (see FIG. 4). In this modification, the second bottom portion 33 has a curved portion overlapping the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction). The lower surface of the second bottom portion 33 is in contact with the thin portion 238 . Note that the thin portion 238 may have a curved shape in its natural state, but is not limited to this. For example, the thin portion 238 may be flat in its natural state, and may be curved when covered with the second bottom portion 33 of the cover member 3 .
　本変形例の細胞培養容器Ａ１２は、培養細胞と培養液を収容し、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。細胞培養容器Ａ１２においては、栓部材２の第１底部２３における天井面２３１は湾曲面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２はポート２４の流路２４２につながっている。このような構成によれば、灌流培養に先立ち容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、当該気泡は湾曲状の天井面２３１を伝って外周縁側に移動する。ここで、ポート２４が上位に位置するように細胞培養容器Ａ１２を傾けると、容器体１内の気泡は、天井面２３１の外周縁付近を伝ってポート２４付近に移動する。そして、当該気泡は、天井凹部２３２を通じて流路２４２内を浮上する。これにより、容器体１内に残存する気泡を細胞培養容器Ａ１２の外部に排出することができる。 The cell culture container A12 of this modified example is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time. In the cell culture vessel A12, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is curved. A ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the air bubbles remain on the curved ceiling surface 231. , and move to the outer edge side. Here, when the cell culture vessel A12 is tilted so that the port 24 is positioned at the top, air bubbles in the container body 1 move along the periphery of the ceiling surface 231 to the vicinity of the port 24 . Then, the air bubbles float in the channel 242 through the ceiling recess 232 . Thereby, the air bubbles remaining in the container body 1 can be discharged to the outside of the cell culture container A12.
　その他にも、本変形例の細胞培養容器Ａ１２において、上記の細胞培養容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A12 of this modified example has the same effects as the cell culture vessel A1 described above.
＜第１実施形態の第３変形例＞
　図１４は、第１実施形態の第３変形例に係る細胞培養容器を示している。本変形例の細胞培養容器Ａ１３においては、一対のポート２４のうち片方の構成が上記実施形態の細胞培養容器Ａ１と異なっている。本変形例において、図１４の左側に位置するポート２４は上記実施形態と同様であるが、図中右側に位置するポート２４の構成が異なっている。図１４の右側に位置するポート２４は、第１底部２３よりも下方に延びる突出部２４７を有する。第１底部２３において、突出部２４７の周囲には天井凹部２３２が形成されていない。 <Third Modification of First Embodiment>
FIG. 14 shows a cell culture vessel according to a third modified example of the first embodiment. In the cell culture vessel A13 of this modified example, the configuration of one of the pair of ports 24 is different from that of the cell culture vessel A1 of the above embodiment. In this modification, the port 24 located on the left side of FIG. 14 is the same as that of the above embodiment, but the configuration of the port 24 located on the right side of the figure is different. The port 24 located on the right side of FIG. 14 has a protrusion 247 extending below the first bottom 23 . The ceiling recess 232 is not formed around the projecting portion 247 in the first bottom portion 23 .
　本変形例の細胞培養容器Ａ１３は、培養細胞と培養液を収容し、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。細胞培養容器Ａ１３においては、栓部材２の第１底部２３における天井面２３１は平坦面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２は、一方のポート２４（図１４における左側）の流路２４２につながっている。このような構成によれば、灌流培養に先立ち容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、一方のポート２４（図１４における左側）が上位に位置するように細胞培養容器Ａ１３を傾けると、容器体１内の気泡は、天井面２３１を伝って上記一方のポート２４付近に移動する。そして、当該気泡は、天井凹部２３２を通じて流路２４２内を浮上する。これにより、容器体１内に残存する気泡を細胞培養容器Ａ１３の外部に排出することができる。そして、灌流培養を行う際には、図１４の右側に位置するポート２４を通じて新しい培養液Ｃが容器体１（細胞培養容器Ａ１３）に供給され、かつ図１４の左側に位置するポート２４を通じて容器体１内（細胞培養容器Ａ１３内部）の培養液が外部に排出される。灌流培養工程においては、容器体１内の培養液を僅かずつ入れ替え続ける。 The cell culture vessel A13 of this modified example is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time. In the cell culture vessel A13, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. The first bottom portion 23 is formed with a ceiling recess 232 recessed upward from the ceiling surface 231 , and the ceiling recess 232 is connected to a flow path 242 of one port 24 (left side in FIG. 14). According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture solution (including cells) prior to perfusion culture, one port 24 (left side in FIG. 14) ) is positioned at the top, air bubbles in the container body 1 move along the ceiling surface 231 to the vicinity of the one port 24 . Then, the air bubbles float in the channel 242 through the ceiling recess 232 . Thereby, the air bubbles remaining in the container body 1 can be discharged to the outside of the cell culture container A13. Then, when perfusion culture is performed, a new culture solution C is supplied to the container body 1 (cell culture container A13) through the port 24 located on the right side of FIG. The culture solution inside the body 1 (inside the cell culture container A13) is discharged to the outside. In the perfusion culture process, the culture solution in the container body 1 is continuously replaced little by little.
　その他にも、本変形例の細胞培養容器Ａ１３において、上記の細胞培養容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A13 of this modified example has the same effects as the cell culture vessel A1 described above.
＜第１実施形態の第４変形例＞
　図１５は、第１実施形態の第４変形例に係る細胞培養容器を示している。本変形例の細胞培養容器Ａ１４においては、カバー部材３の第２底部３３の構成が上記実施形態の細胞培養容器Ａ１と異なっている。本変形例において、第２底部３３には、１つの貫通孔３３１が形成されている。本変形例では、上記実施形態と比べて貫通孔３３１の直径が顕著に大きくされている。貫通孔３３１は、上下方向（第１方向）に見て栓部材２の薄肉部２３８の大半と重なっている。 <Fourth Modification of First Embodiment>
FIG. 15 shows a cell culture vessel according to a fourth modified example of the first embodiment. In the cell culture vessel A14 of this modified example, the configuration of the second bottom portion 33 of the cover member 3 is different from that of the cell culture vessel A1 of the above embodiment. In this modification, one through hole 331 is formed in the second bottom portion 33 . In this modified example, the diameter of the through hole 331 is remarkably increased as compared with the above embodiment. The through hole 331 overlaps most of the thin portion 238 of the plug member 2 when viewed in the vertical direction (first direction).
　本変形例の細胞培養容器Ａ１４は、培養細胞と培養液を収容し、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。細胞培養容器Ａ１４においては、栓部材２の第１底部２３における天井面２３１は平坦面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２はポート２４の流路２４２につながっている。このような構成によれば、灌流培養に先立ち容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、ポート２４が上位に位置するように細胞培養容器Ａ１４を傾けると、容器体１内の気泡は、天井面２３１を伝ってポート２４付近に移動する。そして、当該気泡は、天井凹部２３２を通じて流路２４２内を浮上する。これにより、容器体１内に残存する気泡を細胞培養容器Ａ１４の外部に排出することができる。 The cell culture vessel A14 of this modification is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of culture solution is discharged at the same time. In the cell culture vessel A14, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. A ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position. When the cell culture container A14 is tilted, air bubbles in the container body 1 move along the ceiling surface 231 to the vicinity of the port 24. Then, the air bubbles float in the channel 242 through the ceiling recess 232 . As a result, air bubbles remaining in the vessel body 1 can be discharged to the outside of the cell culture vessel A14.
　その他にも、本変形例の細胞培養容器Ａ１４において、上記の細胞培養容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A14 of this modified example has the same effects as the cell culture vessel A1 described above.
＜第２実施形態＞
　図１６は、本開示の第２実施形態にかかる細胞培養容器を示している。本実施形態の細胞培養容器Ａ２は、ポート２４の構成が上記実施形態の細胞培養容器Ａ１と異なっている。本実施形態においては、ポート２４は、上記実施形態の流路２４２に代えて、第１流路２４３、第２流路２４４および非貫通部２４６を有する。 <Second embodiment>
FIG. 16 shows a cell culture vessel according to a second embodiment of the present disclosure. The cell culture vessel A2 of this embodiment differs from the cell culture vessel A1 of the above embodiment in the configuration of the port 24 . In this embodiment, the port 24 has a first channel 243, a second channel 244, and a non-penetrating portion 246 instead of the channel 242 of the above embodiment.
　上述のように、上記第１実施形態における流路２４２は外部と容器体１の内側空間とに通じる連通路であったが、本実施形態において、第１流路２４３および第２流路２４４は、上下方向（第１方向）において互いに分離している。第１流路２４３は、ポート２４の上部寄りに位置し、外部に開放している。第２流路２４４は、ポート２４の下部寄りに位置し、容器体１の内側空間に開放するとともに天井凹部２３２がつながっている。第１流路２４３および第２流路２４４は、上下方向（第１方向）に見て重なっている。非貫通部２４６は、第１流路２４３と第２流路２４４との間に介在する部分である。 As described above, the flow path 242 in the first embodiment is a communication path that communicates with the outside and the inner space of the container body 1, but in the present embodiment, the first flow path 243 and the second flow path 244 are , are separated from each other in the vertical direction (first direction). The first channel 243 is located near the upper portion of the port 24 and is open to the outside. The second channel 244 is located near the bottom of the port 24 , opens to the inner space of the container body 1 , and is connected to the ceiling recess 232 . The first flow path 243 and the second flow path 244 overlap each other when viewed in the vertical direction (first direction). The non-penetrating portion 246 is a portion interposed between the first channel 243 and the second channel 244 .
　本実施形態の細胞培養容器Ａ２は、培養細胞と培養液を収容し、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。本実施形態においては、容器体１の内側空間への細胞や培養液の導入および廃液の排出は、例えば、容器体１に栓部材２、カバー部材３および外蓋４を装着して細胞培養容器Ａ１をアセンブリ状態とした後に、ポート２４の非貫通部２４６に注射針（図示略）を突き刺して行う。ここで、当該注射針の先端は、非貫通部２４６の直下の第２流路２４４に配置される。 The cell culture vessel A2 of the present embodiment is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of the culture solution is discharged at the same time. In this embodiment, the introduction of cells and culture solution into the inner space of the container body 1 and the discharge of the waste liquid are performed, for example, by attaching the plug member 2, the cover member 3 and the outer lid 4 to the container body 1, and After A1 is brought into an assembled state, the non-penetration portion 246 of the port 24 is pierced with an injection needle (not shown). Here, the tip of the injection needle is arranged in the second flow path 244 immediately below the non-penetrating portion 246 .
　細胞培養容器Ａ２においては、栓部材２の第１底部２３における天井面２３１は平坦面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２はポート２４の流路２４２につながっている。このような構成によれば、このような構成によれば、灌流培養に先立ち容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、ポート２４が上位に位置するように細胞培養容器Ａ２を傾けると、容器体１内の気泡は、天井面２３１を伝ってポート２４付近に移動する。そして、当該気泡は、天井凹部２３２を通じて第２流路２４４内に移動し、上記注射針を介して排出させられる。これにより、容器体１内に残存する気泡を細胞培養容器Ａ２の外部に排出することができる。 In the cell culture vessel A2, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is a flat surface. A ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port When the cell culture vessel A2 is tilted so that the port 24 is positioned at the top, air bubbles in the vessel body 1 move along the ceiling surface 231 to the vicinity of the port 24 . Then, the air bubbles move into the second channel 244 through the recessed ceiling 232 and are discharged through the injection needle. As a result, air bubbles remaining in the container body 1 can be discharged to the outside of the cell culture container A2.
　栓部材２に設けられたポート２４は、第１底部２３から上方に延びる延出部２４１を有する。また、カバー部材３は第５筒状部３４を含み、第５筒状部３４は、延出部２４１の外周面２４１ａに当接して当該外周面２４１ａを覆う。このような構成によれば、ポート２４（非貫通部２４６）に注射針を刺した場合、延出部２４１が径方向外方に膨らむことは、第５筒状部３４によって抑制される。また、注射針を非貫通部２４６から抜いた後、注射針による孔が閉じる。上記構成のポート２４（非貫通部２４６）は、注射針の突き刺し孔を塞ぐ再シール性を有する。 The port 24 provided on the plug member 2 has an extending portion 241 extending upward from the first bottom portion 23 . Further, the cover member 3 includes a fifth cylindrical portion 34, and the fifth cylindrical portion 34 contacts the outer peripheral surface 241a of the extending portion 241 to cover the outer peripheral surface 241a. According to such a configuration, when the port 24 (non-penetration portion 246) is pierced with an injection needle, the extension portion 241 is prevented from expanding radially outward by the fifth tubular portion 34. As shown in FIG. Also, after the injection needle is pulled out from the non-penetration portion 246, the injection needle hole is closed. The port 24 (non-penetration portion 246) configured as described above has a resealing property to block the puncture hole of the injection needle.
　その他にも、本実施形態の細胞培養容器Ａ２において、上記の細胞培養容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A2 of this embodiment has the same effects as the cell culture vessel A1 described above.
＜第３実施形態＞
　図１７は、本開示の第３実施形態にかかる細胞培養容器を示している。本実施形態の細胞培養容器Ａ３は、上記第１実施形態の細胞培養容器Ａ１と異なり、カバー部材３を具備していない。即ち、細胞培養容器Ａ３は、容器体１、栓部材２および外蓋４を備えている。 <Third Embodiment>
FIG. 17 shows a cell culture vessel according to the third embodiment of the present disclosure. The cell culture vessel A3 of this embodiment does not include the cover member 3 unlike the cell culture vessel A1 of the first embodiment. That is, the cell culture vessel A3 has a vessel body 1, a plug member 2 and an outer lid 4. As shown in FIG.
　細胞培養容器Ａ３において、容器体１および外蓋４の構成は上記第１実施形態と同様である。その一方、栓部材２の構成が上記第１実施形態と大きく異なる。 In the cell culture vessel A3, the configurations of the vessel body 1 and the outer lid 4 are the same as in the first embodiment. On the other hand, the configuration of the plug member 2 is significantly different from that of the first embodiment.
　本実施形態において、栓部材２は、第１部２Ａおよび第２部２Ｂを備えて構成される。第２部２Ｂは、可撓性、弾力性を有する軟質素材からなり、第２筒状部２１および第１フランジ部２２を有する。第２部２Ｂは、第１部２Ａと容器体１の第１筒状部１１との間に介在している。本実施形態では、上記第１実施形態と同様に、第２筒状部２１の外周部には一対の環状突起２１１が上下方向（第１方向）に間隔を隔てて形成されている。容器体１への栓部材２（第１部２Ａおよび第２部２Ｂ）の装着時には、第２筒状部２１は、径方向内方に圧縮されつつ第１筒状部１１に内嵌される。そして環状突起２１１が第１筒状部１１の内周面に密着して密閉する。第２部２Ｂを構成する素材としては、例えば、シリコーンゴム、エラストマー樹脂などが挙げられる。 In this embodiment, the plug member 2 is configured with a first portion 2A and a second portion 2B. The second part 2B is made of a soft material having flexibility and elasticity, and has a second tubular part 21 and a first flange part 22 . The second portion 2B is interposed between the first portion 2A and the first cylindrical portion 11 of the container body 1. As shown in FIG. In this embodiment, as in the first embodiment, a pair of annular projections 211 are formed on the outer peripheral portion of the second cylindrical portion 21 with a gap therebetween in the vertical direction (first direction). When the plug member 2 (the first portion 2A and the second portion 2B) is attached to the container body 1, the second tubular portion 21 is radially inwardly compressed and fitted into the first tubular portion 11. . Then, the annular protrusion 211 is in close contact with the inner peripheral surface of the first cylindrical portion 11 to seal it. Examples of the material forming the second portion 2B include silicone rubber and elastomer resin.
　第１部２Ａは、第１底部２３、ポート２４、内側筒状部２６および上側フランジ部２７を含む。第１底部２３は、略一定の厚さとされている。これにより、本実施形態では、上記第１実施形態と異なり、第１底部２３は薄肉部２３８を有さない。第１底部２３に形成された天井凹部２３２は、錐形状部２３３および周縁部２３４を有する。錐形状部２３３は、流路２４２につながっている。図示した例では、錐形状部２３３は、流路２４２とつながる位置から下方に向かうにつれて横断面の面積が大きくなるように形成された円錐形状部分である。周縁部２３４は、錐形状部２３３につながり、かつ第１底部２３の外周縁に通じている。周縁部２３４は、第１底部２３の径方向外方に向かうにつれて下方に位置するように傾斜している。内側筒状部２６は、第１底部２３の外周縁から上方に延びており、概略円筒状とされている。内側筒状部２６は、第２筒状部２１の内周面を覆っている。 The first portion 2A includes a first bottom portion 23, a port 24, an inner cylindrical portion 26 and an upper flange portion 27. The first bottom portion 23 has a substantially constant thickness. Accordingly, in this embodiment, unlike the first embodiment, the first bottom portion 23 does not have the thin portion 238 . A ceiling recess 232 formed in the first bottom portion 23 has a conical portion 233 and a peripheral portion 234 . The conical portion 233 is connected to the channel 242 . In the illustrated example, the cone-shaped portion 233 is a cone-shaped portion formed so that the cross-sectional area increases downward from the position where it connects with the flow path 242 . The peripheral edge portion 234 is connected to the conical portion 233 and communicates with the outer peripheral edge of the first bottom portion 23 . The peripheral edge portion 234 is inclined downward as it goes radially outward of the first bottom portion 23 . The inner tubular portion 26 extends upward from the outer peripheral edge of the first bottom portion 23 and has a substantially cylindrical shape. The inner tubular portion 26 covers the inner peripheral surface of the second tubular portion 21 .
　上側フランジ部２７は、概略円環状とされており、内側筒状部２６の上端から径方向外方に延出している。上側フランジ部２７は、当該上側フランジ部２７の厚さ方向に見て第１フランジ部２２および第１筒状部１１と重なっている。 The upper flange portion 27 has a substantially annular shape and extends radially outward from the upper end of the inner cylindrical portion 26 . The upper flange portion 27 overlaps the first flange portion 22 and the first tubular portion 11 when viewed in the thickness direction of the upper flange portion 27 .
　第１部２Ａは、硬質素材からなり、例えば半透明または透明のプラスチック材料により形成されている。当該プラスチック材料としては、例えばポリスチレンやメチルペンテンのほか、ポリカーボネート、シクロオレフィンポリマー、シクロオレフィンコポリマーなどの、好適には透明性を有する材料が用いられるが、これらに限定されない。 The first part 2A is made of a hard material, such as a translucent or transparent plastic material. Examples of the plastic material include, but are not limited to, polystyrene, methylpentene, polycarbonate, cycloolefin polymer, cycloolefin copolymer, and other transparent materials.
　本実施形態の細胞培養容器Ａ３は、培養細胞と培養液を収容し、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。細胞培養容器Ａ３においては、栓部材２の第１底部２３における天井面２３１は平坦面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２はポート２４の流路２４２につながっている。このような構成によれば、灌流培養に先立ち容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、ポート２４が上位に位置するように細胞培養容器Ａ３を傾けると、容器体１内の気泡は、天井面２３１を伝ってポート２４付近に移動する。そして、当該気泡は、天井凹部２３２を通じて流路２４２内を浮上する。これにより、容器体１内に残存する気泡を細胞培養容器Ａ１１の外部に排出することができる。 The cell culture vessel A3 of the present embodiment is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of the culture solution is discharged at the same time. In the cell culture vessel A3, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. A ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position. When the cell culture container A3 is tilted, air bubbles in the container body 1 move along the ceiling surface 231 to the vicinity of the port 24. FIG. Then, the air bubbles float in the channel 242 through the ceiling recess 232 . As a result, air bubbles remaining in the vessel body 1 can be discharged to the outside of the cell culture vessel A11.
　本実施形態では、天井凹部２３２（周縁部２３４）は、第１底部２３の外周縁に通じている。このような構成によれば、容器体１の内側空間に残存する気泡を排出するために細胞培養容器Ａ３を傾けた際、気泡が第１底部２３の外周縁ないし第２筒状部２１の外周縁側に到達しても、当該気泡を、周縁部２３４を伝わせて流路２４２に導くことができる。これにより、容器体１の内側空間に残存する気泡を、より的確に排出することが可能である。 In this embodiment, the ceiling recessed portion 232 (peripheral edge portion 234) communicates with the outer peripheral edge of the first bottom portion 23. According to such a configuration, when the cell culture container A3 is tilted in order to discharge the air bubbles remaining in the inner space of the container body 1, the air bubbles are removed from the outer peripheral edge of the first bottom portion 23 or the outer peripheral edge of the second cylindrical portion 21. Even if it reaches the edge side, the air bubble can be guided to the channel 242 along the peripheral edge portion 234 . Thereby, it is possible to more accurately discharge the air bubbles remaining in the inner space of the container body 1 .
　その他にも、本実施形態の細胞培養容器Ａ３は、図１～図９等を参照して説明した上記第１実施形態の細胞培養容器Ａ１と同様の構成の範囲において、上記の細胞培養容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A3 of the present embodiment has the same structure as the cell culture vessel A1 of the first embodiment described with reference to FIGS. It has the same function and effect as.
＜第４実施形態＞
　図１８は、本開示の第４実施形態にかかる細胞培養容器を示している。本実施形態の細胞培養容器Ａ４は、上記第１実施形態の細胞培養容器Ａ１と異なり、カバー部材３および外蓋４を具備していない。即ち、細胞培養容器Ａ４は、容器体１および栓部材２を備えている。 <Fourth Embodiment>
FIG. 18 shows a cell culture vessel according to a fourth embodiment of the present disclosure. Unlike the cell culture vessel A1 of the first embodiment, the cell culture vessel A4 of this embodiment does not have the cover member 3 and the outer lid 4. As shown in FIG. That is, the cell culture vessel A4 has a container body 1 and a plug member 2. As shown in FIG.
　本実施形態では、外蓋４を具備しないことに伴い、容器体１における第１筒状部１１の外周面には雄ネジ１１１が形成されていない。 In this embodiment, since the outer lid 4 is not provided, the external thread 111 is not formed on the outer peripheral surface of the first cylindrical portion 11 of the container body 1 .
　栓部材２は、例えばゴム成形品であり、第２筒状部２１、第１フランジ部２２、第１底部２３および一対のポート２４を含む。栓部材２は、可撓性、弾力性を有する軟質素材からなる。栓部材２を構成する素材は、上記第１実施形態と同様である。本実施形態において、第２筒状部２１の外周部には環状突起２１１が形成されていない。第２筒状部２１の外径寸法は、自然状態において第１筒状部１１の内径寸法よりも大きくされている。上記構成の第２筒状部２１は、第１筒状部１１に圧入される。これにより、第２筒状部２１の外周面が第１筒状部１１の内周面に面的に密着（当接）して密閉する。また、本実施形態において、第１底部２３は、薄肉部２３８を有さない。第１底部２３に形成された天井凹部２３２は、錐形状とされており、第１底部２３からポート２４（延出部２４１）に跨って形成されている。 The plug member 2 is, for example, a molded rubber product, and includes a second cylindrical portion 21, a first flange portion 22, a first bottom portion 23 and a pair of ports 24. The plug member 2 is made of a soft material having flexibility and elasticity. The material constituting the plug member 2 is the same as that of the first embodiment. In this embodiment, the annular protrusion 211 is not formed on the outer peripheral portion of the second tubular portion 21 . The outer diameter dimension of the second tubular portion 21 is larger than the inner diameter dimension of the first tubular portion 11 in the natural state. The second tubular portion 21 configured as described above is press-fitted into the first tubular portion 11 . As a result, the outer peripheral surface of the second cylindrical portion 21 is in close contact with (abuts) the inner peripheral surface of the first cylindrical portion 11 to form a tight seal. Also, in the present embodiment, the first bottom portion 23 does not have the thin portion 238 . The ceiling concave portion 232 formed in the first bottom portion 23 has a conical shape and is formed across the port 24 (extending portion 241) from the first bottom portion 23. As shown in FIG.
　本実施形態の細胞培養容器Ａ４は、培養細胞と培養液を収容し、自動的に一定の低速度で常時培養液を供給し、同時に同量の培養液を排出する灌流培養に使用される。細胞培養容器Ａ４においては、栓部材２の第１底部２３における天井面２３１は平坦面とされている。第１底部２３には、この天井面２３１から上方に凹む天井凹部２３２が形成されており、当該天井凹部２３２はポート２４の流路２４２につながっている。このような構成によれば、灌流培養に先立ち容器体１の内側空間を培養液（細胞を含む）で満たす際に当該内側空間に気泡が残存しても、ポート２４が上位に位置するように細胞培養容器Ａ４を傾けると、容器体１内の気泡は、天井面２３１を伝ってポート２４付近に移動する。そして、当該気泡は、天井凹部２３２を通じて流路２４２内を浮上する。これにより、容器体１内に残存する気泡を細胞培養容器Ａ４の外部に排出することができる。 The cell culture vessel A4 of this embodiment is used for perfusion culture in which cultured cells and a culture solution are accommodated, the culture solution is automatically supplied at a constant low speed, and the same amount of the culture solution is discharged at the same time. In the cell culture vessel A4, the ceiling surface 231 of the first bottom portion 23 of the plug member 2 is flat. A ceiling recess 232 that is recessed upward from the ceiling surface 231 is formed in the first bottom portion 23 , and the ceiling recess 232 is connected to the flow path 242 of the port 24 . According to such a configuration, even if air bubbles remain in the inner space when the inner space of the container body 1 is filled with the culture medium (including cells) prior to perfusion culture, the port 24 is positioned at the upper position. When the cell culture container A4 is tilted, air bubbles in the container body 1 move along the ceiling surface 231 to the vicinity of the port 24. FIG. Then, the air bubbles float in the channel 242 through the ceiling recess 232 . As a result, air bubbles remaining in the container body 1 can be discharged to the outside of the cell culture container A4.
　その他にも、本実施形態の細胞培養容器Ａ４は、図１～図９等を参照して説明した上記第１実施形態の細胞培養容器Ａ１と同様の構成の範囲において、上記の細胞培養容器Ａ１と同様の作用効果を奏する。 In addition, the cell culture vessel A4 of the present embodiment has the same structure as the cell culture vessel A1 of the first embodiment described with reference to FIGS. It has the same function and effect as.
　以上、本開示の具体的な実施形態を説明したが、本開示にかかる細胞培養容器は、上記実施形態に限定されるものではない。本開示の細胞培養容器の各部の具体的な構成は、種々に設計変更自在である。 Although specific embodiments of the present disclosure have been described above, the cell culture vessel according to the present disclosure is not limited to the above embodiments. The specific configuration of each part of the cell culture vessel of the present disclosure can be changed in various ways.
　Ａ１，Ａ１１，Ａ１２，Ａ１３，Ａ１４，Ａ２，Ａ３，Ａ４：細胞培養容器、Ｂ：気泡、Ｃ：培養液、１：容器体、１１：第１筒状部、１１０：開口、１１１：雄ネジ（係止手段）、１１２：スリット、１１３：延出片、１１４：上端（第１筒状部の第１方向一方側端）、１２：容器底部、１２１：スカート部、２：栓部材、２Ａ：第１部、２Ｂ：第２部、２１：第２筒状部、２１１：環状突起、２２：第１フランジ部、２３：第１底部、２３１：天井面、２３２：天井凹部、２３３：錐形状部、２３４：周縁部、２３５：外周側厚肉部、２３７：外周縁、２３８：薄肉部、２４：ポート、２４１：延出部、２４１ａ：外周面、２４２：流路、２４３：第１流路、２４４：第２流路、２４５：突起部、２４６：非貫通部、２４７：突出部、２６：内側筒状部、２７：上側フランジ部、３：カバー部材、３１：第２フランジ部、３２：第４筒状部、３３：第２底部、３３１：貫通孔、３４：第５筒状部、４：外蓋、４１：頂板部、４１ａ：主面、４１１：貫通孔、４１２：凸状部、４１２ａ：先端、４２：第３筒状部、４２１：雌ネジ（係止手段）、７：コネクタ、８：チューブ A1, A11, A12, A13, A14, A2, A3, A4: cell culture vessel, B: air bubble, C: culture solution, 1: container body, 11: first cylindrical portion, 110: opening, 111: male screw (locking means), 112: slit, 113: extension piece, 114: upper end (first direction one side end of the first cylindrical portion), 12: container bottom, 121: skirt portion, 2: plug member, 2A : first part, 2B: second part, 21: second tubular part, 211: annular projection, 22: first flange part, 23: first bottom part, 231: ceiling surface, 232: ceiling concave part, 233: cone shape portion 234: peripheral portion 235: outer peripheral side thick portion 237: outer peripheral edge 238: thin portion 24: port 241: extension portion 241a: outer peripheral surface 242: flow path 243: first Flow path 244: Second flow path 245: Protruding portion 246: Non-penetrating portion 247: Protruding portion 26: Inner cylindrical portion 27: Upper flange portion 3: Cover member 31: Second flange portion , 32: fourth cylindrical portion, 33: second bottom portion, 331: through hole, 34: fifth cylindrical portion, 4: outer lid, 41: top plate portion, 41a: main surface, 411: through hole, 412: Convex portion 412a: tip 42: third cylindrical portion 421: female thread (locking means) 7: connector 8: tube

Claims (11)

  1. 　第１方向の一方側端に開口を有し、かつ前記第１方向に延びる第１筒状部を有する容器体と、
    　前記開口を閉塞可能な栓部材と、を備え、
    　前記栓部材は、前記第１筒状部の内周面に当接して密閉する第２筒状部、前記第２筒状部の前記第１方向における前記一方側端から径方向外方に延出し、前記第１筒状部の前記第１方向における前記一方側端を塞ぐ第１フランジ部、および前記第２筒状部の前記第１方向における他方側端から径方向内方に延出し、前記第１方向に見て前記第２筒状部の内側を塞ぐ板状の第１底部、を含み、
    　前記第１底部には、前記第１方向の前記一方側端が外部に開放し、かつ前記第１方向の前記他方側端が前記容器体の内側空間において開放する流路を各々が有する複数のポートが設けられており、
    　前記第１底部は、前記第１方向の他方側を向く天井面を有し、
    　前記第１底部には、前記天井面から前記第１方向の前記一方側に凹む天井凹部が形成されており、
    　前記天井凹部は、前記複数のポートのうち少なくともいずれか１つにおける前記流路につながっている、細胞培養容器。     a container body having an opening at one end in a first direction and having a first cylindrical portion extending in the first direction;
    a plug member capable of closing the opening,
    The plug member includes a second cylindrical portion that abuts and seals the inner peripheral surface of the first cylindrical portion, and extends radially outward from the one side end of the second cylindrical portion in the first direction. extending radially inward from a first flange portion that closes the one side end of the first tubular portion in the first direction and the other side end of the second tubular portion in the first direction; including a plate-shaped first bottom portion that closes the inside of the second tubular portion when viewed in the first direction,
    In the first bottom portion, a plurality of flow paths each having a flow path whose one side end in the first direction is open to the outside and the other side end in the first direction is open to the inner space of the container body A port is provided
    The first bottom has a ceiling surface facing the other side in the first direction,
    The first bottom portion is formed with a ceiling recess that is recessed from the ceiling surface toward the one side in the first direction,
    The cell culture vessel, wherein the ceiling recess is connected to the flow channel in at least one of the plurality of ports.
  2. 　前記天井凹部は、前記第１底部の外周縁に通じる、請求項１に記載の細胞培養容器。 The cell culture vessel according to claim 1, wherein the ceiling recess communicates with the outer peripheral edge of the first bottom.
  3. 　環状の頂板部と、この頂板部の外周縁から当該頂板部の厚さ方向に延びており、前記第１筒状部に外嵌される第３筒状部と、を有する外蓋をさらに備え、
    　前記容器体および前記外蓋の少なくともいずれか一方には、前記第１筒状部に前記第１フランジ部が当接する状態で前記第１筒状部と前記第３筒状部との相対移動を阻止する係止手段が設けられている、請求項１または２に記載の細胞培養容器。     An outer lid having an annular top plate portion and a third tubular portion extending from the outer peripheral edge of the top plate portion in the thickness direction of the top plate portion and fitted onto the first tubular portion is further provided. ,
    Relative movement between the first tubular portion and the third tubular portion is provided in at least one of the container body and the outer lid while the first flange portion is in contact with the first tubular portion. 3. The cell culture vessel according to claim 1 or 2, further comprising locking means for blocking.
  4. 　前記栓部材は、軟質素材からなる、請求項１ないし３のいずれかに記載の細胞培養容器。 The cell culture vessel according to any one of claims 1 to 3, wherein the plug member is made of a soft material.
  5. 　硬質素材からなり、前記栓部材に重なって当接するカバー部材をさらに備える、請求項４に記載の細胞培養容器。 The cell culture vessel according to claim 4, further comprising a cover member made of a hard material and overlapping and abutting on the plug member.
  6. 　前記カバー部材は、前記第１フランジ部を覆う第２フランジ部と、前記第２筒状部の内周面を覆う第４筒状部と、前記第１底部の少なくとも一部を覆う第２底部と、を含む、請求項５に記載の細胞培養容器。 The cover member includes a second flange portion covering the first flange portion, a fourth tubular portion covering an inner peripheral surface of the second tubular portion, and a second bottom portion covering at least a portion of the first bottom portion. And, the cell culture vessel according to claim 5, comprising.
  7. 　前記ポートは、前記第１底部から前記第１方向の前記一方側に延びる延出部を有し、
    　前記カバー部材は、前記延出部の外周面に当接して当該外周面を囲う第５筒状部を含む、請求項５または６に記載の細胞培養容器。     the port has an extension extending from the first bottom to the one side in the first direction;
    The cell culture vessel according to claim 5 or 6, wherein the cover member includes a fifth tubular portion that abuts on and surrounds the outer peripheral surface of the extending portion.
  8. 　前記第１底部は、他の部位よりも相対的に厚さが小とされ、かつガス透過性を有する薄肉部を含む、請求項４に記載の細胞培養容器。 The cell culture vessel according to claim 4, wherein the first bottom part has a thickness relatively smaller than that of other parts and includes a gas-permeable thin-walled part.
  9. 　前記第１底部は、他の部位よりも相対的に厚さが小とされ、かつガス透過性を有する薄肉部を含み、
    　前記第２底部には、前記第１方向に貫通する貫通孔が形成されており、
    　前記貫通孔は、前記第１方向に見て前記薄肉部と重なっている、請求項６に記載の細胞培養容器。     The first bottom portion includes a thin portion having a relatively smaller thickness than other portions and having gas permeability,
    A through hole penetrating in the first direction is formed in the second bottom,
    The cell culture vessel according to claim 6, wherein the through-hole overlaps with the thin portion when viewed in the first direction.
  10. 　前記延出部の先端には、前記外周面よりも外向きに突出し、かつ前記第５筒状部の前記第１方向における前記一方側端に当接し得る突起部が設けられている、請求項７に記載の細胞培養容器。 The tip of the extending portion is provided with a protruding portion that protrudes outward from the outer peripheral surface and can come into contact with the one side end of the fifth tubular portion in the first direction. 7. The cell culture vessel according to 7.
  11. 　前記流路は、外部に開放する第１流路、および前記容器体の前記内側空間に開放し、かつ前記天井凹部がつながる第２流路からなり、
    　前記第１流路および前記第２流路は、前記第１方向に互いに離間し、かつ前記第１方向に見て重なっている、請求項７または１０に記載の細胞培養容器。     The flow path comprises a first flow path open to the outside and a second flow path open to the inner space of the container body and connected to the ceiling recess,
    The cell culture vessel according to claim 7 or 10, wherein the first channel and the second channel are separated from each other in the first direction and overlap when viewed in the first direction.
PCT/JP2021/009546 2021-03-10 2021-03-10 Cell culture vessel WO2022190261A1 (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013543738A (en) * 2010-11-22 2013-12-09 コーニング インコーポレイテッド Hermetic assembly for cell culture equipment
WO2019082261A1 (en) * 2017-10-24 2019-05-02 株式会社サンプラテック Cell container
WO2021014642A1 (en) * 2019-07-25 2021-01-28 株式会社サンプラテック Container for housing culture solution

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013543738A (en) * 2010-11-22 2013-12-09 コーニング インコーポレイテッド Hermetic assembly for cell culture equipment
WO2019082261A1 (en) * 2017-10-24 2019-05-02 株式会社サンプラテック Cell container
WO2021014642A1 (en) * 2019-07-25 2021-01-28 株式会社サンプラテック Container for housing culture solution

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