WO2015059798A1 - Cell or tissue transport device - Google Patents
Cell or tissue transport device Download PDFInfo
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- WO2015059798A1 WO2015059798A1 PCT/JP2013/078855 JP2013078855W WO2015059798A1 WO 2015059798 A1 WO2015059798 A1 WO 2015059798A1 JP 2013078855 W JP2013078855 W JP 2013078855W WO 2015059798 A1 WO2015059798 A1 WO 2015059798A1
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- specific gravity
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- cell
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/52—Mobile; Means for transporting the apparatus
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/20—Material Coatings
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
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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
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/22—Means for packing or storing viable microorganisms
Definitions
- the present invention relates to an apparatus for transporting cells or tissues collected from a living body or produced at a cell processing center.
- CPC cell processing centers
- the present invention has been made in view of the above, and an object of the present invention is to reduce physical impact on cells or tissues during transportation and damage to cell activity, and to easily collect cells after transportation. It is an object of the present invention to provide a device for transporting cells or tissues.
- a feature of the present invention relates to a transport device used for transporting cells or tissues.
- the transport device according to the feature of the present invention is disposed in the holding container so as to be separated from the holding container that stores the high specific gravity solution and the low specific gravity solution in which the interface is formed, and the cell or tissue held in the interface.
- a recovery tub for recovering cells or tissues held at the interface is disposed in the holding container so as to be separated from the holding container that stores the high specific gravity solution and the low specific gravity solution in which the interface is formed, and the cell or tissue held in the interface.
- FIG. 1 is a perspective view showing an appearance of a transportation apparatus according to an embodiment of the present invention.
- FIG. 2 is a schematic view showing a longitudinal section showing the internal structure of the transportation apparatus shown in FIG.
- FIG. 3A is a perspective view showing a configuration of an inner lid used in the transportation apparatus shown in FIG.
- FIG. 3B is a cross-sectional view showing the configuration of the inner lid used in the transportation device shown in FIG. 1.
- FIG. 4 is a perspective view showing a configuration of a centrifuge tube and an inner cap used in a transportation apparatus according to another embodiment of the present invention.
- FIG. 5 is a perspective view showing a state where the centrifuge tube and the inner lid cap shown in FIG. 4 are separated.
- FIG. 6 is a longitudinal sectional view showing a screwed state between the centrifuge tube and the inner lid cap shown in FIG.
- FIG. 7A is a perspective view showing a configuration of an air vent upper stage portion, a conical spreading portion, and a recovery tank provided at an upper portion of the centrifuge tube shown in FIG. 4.
- FIG. 7B is a bottom view showing the configuration of the upper part of the air vent, the conical spreading part, and the recovery tank provided in the upper part of the centrifuge tube shown in FIG. 4.
- FIG. 8A is a front view showing a recovery rod used in the centrifuge tube shown in FIG.
- FIG. 8B is a perspective view showing a recovery rod used in the centrifuge tube shown in FIG. FIG.
- FIG. 9 is a perspective view showing a state where only the centrifuge tube and the inner lid cap shown in FIG. 4 are separated.
- FIG. 10 is a schematic view showing a longitudinal section of a cell storage container used in a transport apparatus according to another embodiment of the present invention.
- FIG. 1 is a perspective view showing an external appearance of a cell or tissue transport apparatus according to an embodiment of the present invention.
- the transport device 1 shown in the figure is configured by being entirely surrounded by a cylindrical and large-diameter outer container 3 made of aluminum, stainless steel, or the like.
- the outer lid 31 is closely fitted in the opening at the upper end of the outer container 3 so that the inside of the outer container 3 can be sealed.
- a centrifuge tube 201 which is a cell or tissue holding container is disposed inside the outer container 3.
- the centrifuge tube 201 accommodates the high specific gravity solution and the low specific gravity solution so as to float and hold the cells at the interface formed between the high specific gravity solution and the low specific gravity solution.
- the lower part of the centrifuge tube 201 is formed in a hemispherical shape or a conical shape.
- a spring 33 s is disposed below the centrifuge tube 201. The centrifuge tube 201 is supported by the spring 33s with buffering properties.
- the USB 35b is connected to the middle of the outer container 3 via an electric cable.
- the USB 35b is connected to a later-described control unit inside the transport apparatus 1 via an electric cable, and is connected to an external control apparatus (not shown).
- the USB 35b supplies power and control signals from the control device to the transport device 1 via the USB 35b, and transmits information from the transport device 1 to an external control device via the USB 35b.
- an LED is provided in the outer container 3 slightly above the portion to which the USB 35b is connected. This LED may notify whether or not the transport device 1 is in an operating state.
- FIG. 2 is a schematic diagram showing a longitudinal section showing the internal structure of the transport apparatus 1 shown in FIG.
- the high specific gravity solution 9 and the low specific gravity solution 11 are placed in the centrifuge tube 201.
- An interface 13 is formed between the high specific gravity solution 9 that sinks downward and the low specific gravity solution 11 that floats above the two types of solutions placed in the centrifuge tube 201.
- the cells 15 are held so as to float on the interface 13.
- the transport device 1 is in a state in which no scaffold is provided for the cells 15, a physical impact due to vibrations during transport is not transmitted to the cells 15, and the cell activity is not damaged. It is said.
- the recovery rod 17 is disposed in the high specific gravity solution 9 below the interface 13. Further, the lower end portion of the tube 21 extending vertically downward from above is fixedly attached to the flange frame 19 surrounding the peripheral edge portion of the recovery rod 17.
- the collection basket 17 is made of, for example, a nylon mesh having a mesh size of 5 ⁇ m.
- the collection basket 17 is configured so that the cells 15 suspended and held at the interface 13 above the collection basket 17 can be scooped up and efficiently collected as described later.
- a protein absorption inhibitor is applied to the inside of the cocoon frame 19, and cell adhesion to the collection tub 17 is prevented.
- the inner lid 23 is fitted inside the upper end of the centrifuge tube 201. As shown in an enlarged view in FIGS. 3A and 3B, the inner lid 23 has an outer peripheral surface that is squeezed downward. The squeezed lower end faces the interface 13 above the interface 13 in the low specific gravity solution 11.
- the inner lid 23 has an air discharge part. As shown in FIG. 3A and FIG. 3B, the air discharge portion spreads conically downward from the lower end portion of the air discharge hole formed substantially at the center.
- the air discharge portion has a conical portion 23 b that is close to the inner wall of the centrifuge tube 201 so that the widened opening at the lower end collects air in the centrifuge tube 201.
- a solution injection hole for injecting the solution into the centrifuge tube 201 is formed near the peripheral edge of the conical portion 23b. The upper end of the tube 21 is inserted into the solution injection hole.
- a short air discharge pipe 23 a is inserted into the air discharge hole at the substantially center of the inner lid 23.
- a sealing portion 23c is formed at the upper end of the air discharge pipe 23a.
- the sealing part 23c seals the air discharge pipe 23a using a sealing sealer.
- a sealing portion 21 c is formed at the upper end of the tube 21 that extends through the periphery of the inner lid 23. The sealing part 21c seals the tube 21 using a sealing sealer.
- each of the air discharge pipe 23a and the tube 21 is sealed using the sealing sealer, so that the inner lid 23 injects the solution from the tube 21 into the centrifuge tube 201.
- the air in the centrifuge tube 201 is completely discharged from the air discharge pipe 23a, the air is not allowed to enter the centrifuge tube 201 from the air discharge pipe 23a and the tube 21.
- the sealing sealer is used for sealing, the upper ends of the air discharge pipe 23a and the tube 21 are cut and opened with scissors or the like.
- the film heater 33g, the carbon 33f, the aluminum tape 33e, the aluminum plate 33d, and the acrylic cylinder 33c are arranged from the inside to the outside so as to entirely cover the outer periphery of the centrifuge tube 201 outside the centrifuge tube 201. In the order of description, they are provided in layers. Further, a heat insulating portion 33b is provided between the acrylic cylinder 33c and the inner peripheral surface of the outer container 3 for heat insulation and buffer suppression. The heat insulating portion 33b is made of expanded polystyrene, aluminum beads, or the like. In addition, although the outer peripheral part of the centrifuge tube 201 and the film heater 33g are spaced apart from each other in the drawing, they are in close contact with each other so that the centrifuge tube 201 can be inserted and removed.
- a temperature logger 31a is provided below the outer lid 31 blocking the upper opening of the outer container 3 and above the air discharge pipe 23a.
- the temperature logger 31a is configured to measure and record the temperature.
- a control unit 35 made of, for example, a one-chip microprocessor is provided on the inner peripheral surface of the outer container 3.
- a USB 35b is connected to the control unit 35 via an electric cable.
- a thermostat 35a is attached to the outer periphery of the acrylic cylinder 33c. The thermostat 35a is connected to the control unit 35 via an electric cable.
- the cells 15 collected from the patient in the medical institution for transport to the CPC are the high specific gravity solution 9 and the low specific gravity solution injected into the centrifuge tube 201. 11 is held so as to float on the interface 13 formed between them.
- the high specific gravity solution 9 and the low specific gravity solution 11 are injected into the centrifuge tube 201 as described above, and the cells 15 are suspended and held at the interface 13 between them. Thereafter, in order to discharge the air in the centrifuge tube 201, the high specific gravity solution 9 or the low specific gravity solution 11 is additionally injected into the centrifuge tube 201 from the tube 21, and the air in the centrifuge tube 201 is changed into the conical portion 23b. Collected in The collected air is discharged to the outside from the air discharge pipe 23a, and the air in the centrifuge tube 201 is discharged. Then, the inner lid 23 is fitted in an airtight state in the upper opening of the centrifuge tube 201, and the upper ends of the tube 21 and the air discharge pipe 23a are sealed with a sealing sealer.
- the cells 15 are held in a suspended state at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11, and air is evacuated from the centrifuge tube 201.
- the centrifuge tube 201 is disposed in the outer container 3. Further, the lower end portion of the centrifuge tube 201 is placed on the spring 33s.
- the control unit 35 drives the film heater 33g to generate heat. Due to this heat generation, the high specific gravity solution 9 and the low specific gravity solution 11 in the centrifuge tube 201 are heated to a predetermined temperature, for example, a temperature of 25 ° C. to 37 ° C., for example, and the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11. The cell 15 held in is warmed to the predetermined temperature. Then, the control unit 35 controls to keep the cells 15 at a predetermined temperature at all times, and keeps the survival rate of the cells 15 high.
- a predetermined temperature for example, a temperature of 25 ° C. to 37 ° C.
- the transport device 1 floats and holds the cells 15 on the interface 13 in the centrifuge tube 201 and keeps the cells 15 at a predetermined temperature, for example, by courier or the like.
- the transport device 1 of this embodiment has an extremely compact and simple structure in which the centrifuge tube 201 containing the high specific gravity solution 9 and the low specific gravity solution 11 is housed in the outer container 3, and is extremely easy to handle in transportation. Is. In this transportation, since the cells 15 are suspended and held at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 in the airtight centrifuge tube 201 from which the air has been exhausted, Impacts such as vibration and damage to cell activity can be reduced.
- the transport device 1 of the present embodiment does not provide a scaffold, it is possible to prevent environmental deterioration in the cell holding container due to cell proliferation by stopping or reducing cell proliferation activity, and at the same time, dormant cells. Can be kept in a state.
- the cells 15 are always maintained at a predetermined optimum temperature by the heating control under the control of the control unit 35, the cells 15 can be transported in a state in which the cell activity is maintained without causing a low-temperature injury as in the prior art. it can.
- the transport device 1 After the transport device 1 is transported to the CPC, it is necessary to collect the cells 15 from the cell transport device 1 in order to produce tissue from the cells 15 in the CPC.
- the outer lid 31 of the outer container 3 of the cell transport device 1 is removed, and the centrifuge tube 201 is taken out from the outer container 3.
- the tube 21 fixed to the inner lid 23 is also lifted upward.
- the recovery rod 17 fixed to the lower end portion of the tube 21 is also lifted and taken out simultaneously while draining the high specific gravity solution 9 and the low specific gravity solution 11 from the mesh. Therefore, the cells 15 held at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 are easily recovered so as to be scooped up by the recovery rod 17.
- the cells 15 can be recovered very simply by simply lifting the inner lid 23 upward from the centrifuge tube 201.
- PFC perfluorocarbon
- gas control of the low specific gravity solution that is, control by gas movement between the PFC and the culture medium or the storage solution is performed. Can do.
- the transport apparatus 1 makes it possible to reduce damage to cell activity during transport by using normal culture conditions such as temperature and pH.
- FIG. 4 and 5 are perspective views showing a centrifuge tube 201 used in a cell or tissue transport apparatus according to another embodiment of the present invention.
- FIG. 5 illustrates a state where the lower centrifuge tube 201 is separated from the upper portion thereof.
- the centrifuge tube 201 shown in FIG. 4 and FIG. 5 contains the high specific gravity solution 9 and the low specific gravity solution 11 therein.
- an interface 13 is formed as shown in FIG. 4 between the high specific gravity solution 9 sinking downward and the low specific gravity solution 11 floating above.
- the cells 15 are suspended and held on the interface 13 and form a state where there is no scaffold for the cells 15. Thereby, the physical impact by the vibration at the time of transport etc. is not transmitted with respect to the cell 15, and it is comprised so that a cell activity may not be damaged.
- a screw portion 201 a is formed outside the upper end portion of the centrifuge tube 201.
- a screw portion 123e formed on the inner side of the inner lid cap 123d fixedly attached and fixed to the inner lid 213 tightly closing the upper opening of the centrifuge tube 201 with an adhesive or the like is attached to the screw portion 201a. Screw together.
- FIG. 6 illustrates this screwed state.
- a conical spreading portion 225 is formed so as to spread in a conical shape downward from a lower end portion of a cylindrical portion 225a formed at the center of the inner lid 213. This widened opening at the lower end collects air in the centrifuge tube 201.
- an air vent pipe 123a is attached through the inner lid 213 and the central portion of the inner lid cap 123d fixed to the inner lid 213.
- the air vent pipe 123a is tightly closed by tightly fitting the air vent cap 123c to the upper end of the air vent pipe 123a.
- the upper part of the air vent pipe 123a to which this inner lid cap 123d is fitted may be configured to be closed by a syringe instead of the air vent cap 123c.
- the lower end portion of the air vent pipe 123a is connected to the upper end portion of the cylindrical portion 225a above the conical spreading portion 225 in the inner lid 213. Air or a solution can pass through both of them, that is, through the air vent pipe 123a and the cylindrical portion 225a.
- the upper end of the tube 121 is provided so as to penetrate near the peripheral edge of the inner lid cap 123d.
- a tube cap 121c is attached to the upper end portion of the tube 121 so as to be tightly fitted.
- the lower part of the tube 121 penetrates the inner lid cap 123d, and further outside the recovery rod 117 described later in the inner lid 213, that is, near the peripheral edge of the conical spreading portion 225 in the inner lid 213. It extends downward through the tube hole 123f formed as shown in FIG.
- the lower end of the tube 121 extends downward into the centrifuge tube 201.
- a collection basket 117 is disposed in the middle of the conical spreading portion 225 of the inner lid 213.
- the collection basket 117 is arranged such that the upper corner bites into the inclined part of the conical spreading part 225.
- a collar frame 119 is provided on the peripheral edge of the collection basket 117.
- the mesh portion of the collection basket 117 is made of, for example, a nylon mesh having a mesh size of 5 ⁇ m.
- the mesh portion of the collection basket 117 is configured so that the collection basket 117 can collect the cells 15 suspended and held at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11.
- a protein absorption inhibitor is applied to the inside of the gutter frame 119 and the centrifuge tube 201, thereby preventing cell adhesion to the collection gutter 117.
- the centrifuge tube 201 of the present embodiment configured as described above holds the cells 15 while floating at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 in the centrifuge tube 201. Furthermore, after the inner lid cap 123d is tightly fitted so that the solution inside does not leak, the centrifuge tube 201 is housed in the transport device 1 and transported to the CPC.
- the CPC that has received the transport device 1 containing the centrifuge tube 201 collects the cell 15 from the centrifuge tube 201 in order to produce tissue from the cell 15.
- the air vent cap 123c and the tube cap 121c are first removed, and a syringe (not shown) is attached to the upper end of the air vent pipe 123a instead of the air vent cap 123c.
- the centrifuge tube 201 is turned upside down, and the high specific gravity solution 9 and the low specific gravity solution 11 are extracted with a syringe attached to the air vent pipe 123a.
- the high specific gravity solution 9 and the low specific gravity solution 11 in the centrifuge tube 201 are extracted, the cells 15 suspended and held at the interface 13 of these solutions are collected so as to ride on the collection basket 117.
- a new centrifuge tube 201 is attached, and the normal state is restored from the upside-down state.
- PBS phosphate buffered saline
- FIG. 10 is a schematic view showing a longitudinal section of a cell storage container used in a cell or tissue transport apparatus according to another embodiment of the present invention.
- the cell storage container 200 shown in the figure is configured such that the upper opening of the container main body 222 and a screw formed inside the upper lid 223d are screwed together to close the container main body 222 in a sealed state.
- a film heater 233f is provided outside the container body 222, and a heat insulating layer 233b is provided outside the film heater 233f. With this configuration, the inside of the container main body 222 can be heated to a predetermined temperature. In addition, the film heater 233f is provided in close contact and is configured integrally with the container.
- a cylindrical portion 311 is provided on the lower surface of the upper lid 223d with its upper end close to it.
- a lower portion of the cylindrical portion 311 is formed as an inner lid 223 of the container main body 222.
- the inner lid 223 forms a conical spreading portion 223b that spreads conically downward from the lower end portion of the cylindrical portion 311.
- the opening at the lower end that expands in a conical shape below the conical expanding portion 223b is in close contact with the inner peripheral wall of the container body 222 so that the inner solution does not leak and is detachable.
- An air vent pipe 311 is attached through the central portion of the inner lid 223.
- An air vent cap 233c is tightly fitted to the upper end of the air vent pipe 311 to close the air vent pipe 311 tightly.
- the upper part of the air vent pipe 311 may be sealed or closed using a sealing sealer shown in FIG. 2 instead of the air vent cap 233c.
- the high specific gravity solution 9 and the low specific gravity solution 11 are placed in the container main body 222 below the conical spreading portion 223b.
- An interface 13 is formed between the high specific gravity solution 9 that sinks downward and the low specific gravity solution 11 that floats above the two types of solutions placed in the container body 222.
- a tissue piece 215 which is a cell sheet such as a skin tissue cultured on the interface 13 is suspended and held. The tissue piece 215 is configured such that no scaffold is formed, the physical shock due to vibration during transportation is not transmitted to the tissue piece 215, and the cell activity is not damaged. .
- the container body 222 is entirely closed with the inner lid 223. Further, the high specific gravity solution 9 and the low specific gravity solution 11 are placed in the container main body 222 whose upper opening is closed in a sealed state by the upper lid 223d, and the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 is placed in the container body 222.
- the tissue piece 215 is held floating. Thereby, the tissue piece 215 can be safely transported without transferring a physical impact due to vibration or the like to the tissue piece 215 even when the cell storage container 200 is transported.
- the cell or tissue transport apparatus 1 of this embodiment is a cell and tissue transport apparatus used when transporting cells or tissues.
- the transport apparatus 1 includes a centrifuge tube 201 (holding container) that accommodates a high density solution and a low specific gravity solution in which an interface is formed, and a cell or tissue held at the interface, and the centrifuge tube 201 (holding container). And a recovery rod 17 for recovering cells or tissue held at the interface.
- Such a transport device 1 holds cells or tissues at the interface formed between the high specific gravity solution and the low specific gravity solution, and disposes a collection basket 17 apart from the cells held at the interface, The cells held at the interface are recovered by the recovery rod 17.
- the transport device 1 of the present embodiment can safely transport cells or tissues without causing physical impact due to vibration during transport to the cells and without damaging the cell activity.
- the transport device 1 of the present embodiment can easily and efficiently collect cells or tissues held at the interface.
- the transport apparatus 1 further includes an inner lid 23 (container lid) for closing the upper opening of the centrifuge tube 201 (holding container), an air discharge unit, and an air discharge pipe so as to seal the centrifuge tube 201 (holding container). 23a and a tube 21 (solution injection pipe).
- the air discharge part has an air discharge hole, a conical part 23b (conical spreading part) and a solution injection hole.
- the air discharge hole is disposed below the inner lid 23 (container lid) and is formed substantially at the center.
- the conical portion 23b (conical spreading portion) spreads conically downward from the lower end of the air discharge hole, and the opening at the lower end of the air collecting collects the air in the centrifuge tube 201 (holding container).
- the centrifuge tube 201 (holding container) is disposed close to the inner wall.
- the solution injection hole is formed near the peripheral edge of the conical portion 23b (conical spreading portion), and is arranged for introducing the solution into the centrifuge tube 201 (holding container).
- the lower end of the air discharge pipe 23a is inserted into the air discharge hole, the upper end protrudes upward, and is closely fitted into a cylindrical central recess formed at the approximate center of the lower surface of the container lid.
- the tube 21 solution injection pipe
- the tube 21 has an upper end that protrudes upward through the solution injection hole and closely fits into a cylindrical peripheral recess formed near the periphery of the lower surface of the inner lid 23 (container lid). It extends downward and holds the collection basket 17 fixedly.
- an air discharge portion is disposed below the inner lid 23 that closes the centrifuge tube 201, and a cone that extends downward in a conical shape from the air discharge hole at the center of the air discharge portion.
- a shaped portion 23b is provided.
- the transport device 1 of the present embodiment allows the air in the centrifuge tube 201 to flow into the centrifuge portion 23b while injecting the solution into the centrifuge tube 201 from the tube 21 that has passed through the solution injection hole near the periphery of the conical portion 23b. Then, air is discharged from the air discharge pipe 23a collected through the air discharge hole.
- the transport apparatus 1 can reduce the shaking of the solution in the centrifuge tube 201 due to vibration during transportation, and can reduce physical impact on cells or tissues. Further, in the transport device 1 of the present embodiment, since the collection basket 17 is fixedly held at the lower end of the tube 21, the collection basket 17 can be easily taken out by lifting the air discharge part together with the tube 21, and the cells can be easily removed. It can be recovered.
- the transport apparatus 1 of the present embodiment is formed on a peripheral wall portion of a centrifuge tube 201 (holding container), and a film heater 33g (heater) for heating the temperature in the centrifuge tube 201 (holding container) to a predetermined temperature.
- the transport device 1 of the present embodiment forms a film heater 33g on the peripheral wall portion of the centrifuge tube 201, and heats the temperature of the centrifuge tube 201 to a predetermined temperature with the film heater 33g.
- the inside of the centrifuge tube 201 is heated to a predetermined temperature, and cells or tissues held at the interface between the high specific gravity solution and the low specific gravity solution are also heated to the predetermined temperature, and the cell survival rate is high. Hold on.
- the high specific gravity solution is perfluorocarbon or modified silicone oil
- the low specific gravity solution is a cell culture medium or physiological saline.
- the transport apparatus 1 includes a high specific gravity solution of perfluorocarbon or modified silicone oil and a low specific gravity solution of a cell culture medium or physiological saline in a centrifuge tube 201, and a high specific gravity solution and a low specific gravity solution Cells are suspended and held at the interface between.
- the transport apparatus 1 of the present embodiment can reduce physical impacts such as vibrations to cells or tissues during transport and damage to cell activity.
- the transport apparatus 1 of this embodiment does not provide a scaffold, it is possible to prevent deterioration of the environment in the centrifuge tube 201 due to cell proliferation by stopping or reducing cell proliferation activity.
- the gist of the transport device 1 of the present embodiment is that the collection basket 17 is configured by a mesh portion made of nylon mesh and a collar frame 119 surrounding the outer peripheral edge of the mesh portion.
- the mesh portion of the collection basket 17 is made of nylon mesh, and the outer peripheral edge of the collection basket 17 is surrounded by the fence frame 119, so that the high specific gravity solution and the low specific gravity solution are removed. In addition, cells or tissues can be recovered efficiently.
- the transport device 1 applies a protein adsorption inhibitor to the inner wall of the frame 119.
- the transport apparatus 1 of the present embodiment applies the protein adsorption inhibitor to the inner wall of the cage frame 19 that surrounds the recovery basket 17, cell adhesion to the recovery basket 17 can be prevented.
- the transport device 1 of the present embodiment floats and holds cells at the interface formed between the high specific gravity solution and the low specific gravity solution, and disposes a recovery basket away from the interface. Collect the cells retained at the interface.
- the transport device 1 according to the present embodiment is capable of transporting cells safely without causing physical impact due to vibration during transport to the cells, damaging cell activity, and floating at the interface.
- the retained cells can be collected easily and efficiently.
- the transport apparatus 1 of this embodiment arrange
- a conical expanse is provided.
- the transport device 1 of the present embodiment injects the air in the container at the conical spreading part while injecting the solution into the cell holding container from the solution injection pipe passing through the solution injection hole near the peripheral edge of the conical spreading part.
- the collected air is discharged from the air discharge pipe that is passed through the air discharge hole, and the recovery tank is fixedly held at the lower end of the solution injection pipe that is passed through the solution injection hole.
- the transport device 1 of the present embodiment can discharge the air in the cell holding container, reduce the shaking of the solution in the cell holding container due to vibration during transportation, and reduce the physical impact on the cell. it can. Further, in the transport device 1 of the present embodiment, since the recovery tub is fixedly held at the lower end of the solution injection pipe, the recovery tub can be easily taken out by lifting the air discharge portion together with the solution injection pipe, and the cells can be easily removed. It can be recovered.
- the transport device 1 of the present embodiment forms a heater on the peripheral wall portion of the cell holding container, and heats the temperature in the cell holding container to a predetermined temperature with this heater.
- the inside of the cell holding container is heated to a predetermined temperature, and the cells held at the interface between the high specific gravity solution and the low specific gravity solution are also heated to the predetermined temperature, Cell viability can be kept high.
- the transport device 1 of the present embodiment puts a high specific gravity solution such as perfluorocarbon or modified silicone oil and a low specific gravity solution such as a cell culture medium or cell preservation solution (for example, physiological saline) in a cell holding container, Cells are suspended and held at the interface between the high density solution and the low density solution.
- a high specific gravity solution such as perfluorocarbon or modified silicone oil
- a low specific gravity solution such as a cell culture medium or cell preservation solution (for example, physiological saline)
- cells for example, physiological saline
- the transport apparatus 1 of this embodiment can reduce physical impacts such as vibrations to cells during transport and damage to cell activity.
- the transport device 1 of the present embodiment does not provide a scaffold, it is possible to prevent deterioration of the environment in the cell holding container due to cell growth by stopping or reducing the cell growth activity.
- the transport device 1 of the present embodiment removes the high specific gravity solution and the low specific gravity solution because the net-like portion of the recovery basket is made of nylon mesh and the outer peripheral edge of the recovery basket is surrounded by a fence frame.
- the cells can be collected efficiently.
- the transport apparatus 1 of the present embodiment applies the protein adsorption inhibitor to the inner wall of the cage frame surrounding the recovery basket, cell adhesion to the recovery basket can be prevented.
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Abstract
The transport device (1) is used when transporting cells or tissue. The transport device (1) comprises: a centrifuge tube (201) for holding a high specific gravity solution and a low specific gravity solution between which an interface is formed; and a retrieval basket (17), which is disposed inside the centrifuge tube (201) separated from the cells or tissue that is being held at the interface and is for retrieving the cells or tissue that is being held at the interface.
Description
本発明は、生体から採取され、またはセルプロセッシングセンタにて生産された、細胞または組織を輸送する装置に関する。
The present invention relates to an apparatus for transporting cells or tissues collected from a living body or produced at a cell processing center.
現在、日本国内で実施されている再生医療のほとんどは、患者自身の細胞を用いた自家培養移植によるものである。多くの場合、自家培養移植において、細胞は、医療機関において患者から採取され、大学や企業などのセルプロセッシングセンタ(CPC)に輸送される。またこのCPCにおいて、細胞から、目的とする細胞および組織が、生産される。そして生産された細胞および組織は、患者に移植するため再び医療機関へ輸送される。
Currently, most of the regenerative medicine currently being conducted in Japan is based on autologous culture transplantation using the patient's own cells. In many cases, in autologous culture transplantation, cells are collected from patients in medical institutions and transported to cell processing centers (CPC) such as universities and companies. In this CPC, target cells and tissues are produced from the cells. The produced cells and tissues are then transported back to the medical institution for transplantation into the patient.
このような細胞および組織の輸送では、温度およびpHなどの通常の培養条件を再現し、かつ輸送時の細胞への物理的衝撃を軽減する必要がある。また細胞および組織の輸送では、細胞増殖による細胞保持容器内の環境悪化を防止することで、細胞活性へのダメージを軽減することに加え、輸送後の細胞の回収を容易にすることが必要である。
In transporting such cells and tissues, it is necessary to reproduce normal culture conditions such as temperature and pH, and to reduce physical impact on the cells during transport. In addition, in transporting cells and tissues, it is necessary to prevent the deterioration of the environment in the cell holding container due to cell growth, thereby reducing damage to cell activity and facilitating recovery of cells after transport. is there.
しかしながら、従来は、上述したような要件を十分に満たしているものはなかった。例えば足場依存性の細胞は、フラスコに接種し、培養面に接着した状態で輸送される。これにより、物理的衝撃が細胞に直接伝わり、細胞活性がダメージを受けるなど、輸送時の細胞への振動影響が生じてしまう問題があった。さらに、制御温度を低温とすることで輸送時の細胞増殖を抑制しているが、増殖以外の細胞活性を維持できなかったり、更に低温状態でのみ温度制御を行い、ガス制御ができないなどの問題があった。
However, conventionally, none of the above-mentioned requirements have been sufficiently satisfied. For example, anchorage-dependent cells are inoculated into a flask and transported while adhering to the culture surface. As a result, there is a problem in that a physical impact is directly transmitted to the cell, and the cell activity is damaged, causing vibration effects on the cell during transportation. In addition, cell growth during transportation is suppressed by lowering the control temperature, but cell activities other than growth cannot be maintained, and problems such as inability to perform gas control by controlling temperature only at low temperatures was there.
このような問題を解決し、上述したような要件を適確に満たしている細胞または組織の輸送装置が要望されている。
There is a demand for a device for transporting cells or tissues that solves such problems and appropriately satisfies the above-described requirements.
本発明は、上記に鑑みてなされたもので、その目的とするところは、輸送時の細胞または組織への物理的衝撃および細胞活性へのダメージを軽減し、輸送後の細胞の回収を容易にすることができる細胞または組織の輸送装置を提供することにある。
The present invention has been made in view of the above, and an object of the present invention is to reduce physical impact on cells or tissues during transportation and damage to cell activity, and to easily collect cells after transportation. It is an object of the present invention to provide a device for transporting cells or tissues.
上記課題を達成するため、本発明の特徴は、細胞または組織を搬送する際に用いられる輸送装置に関する。本発明の特徴に係る輸送装置は、界面が形成される高比重溶液と低比重溶液とを収容する保持容器と、界面に保持される細胞または組織とは離隔して保持容器内に配設され、界面に保持された細胞または組織を回収するための回収笊とを有する。
In order to achieve the above object, a feature of the present invention relates to a transport device used for transporting cells or tissues. The transport device according to the feature of the present invention is disposed in the holding container so as to be separated from the holding container that stores the high specific gravity solution and the low specific gravity solution in which the interface is formed, and the cell or tissue held in the interface. And a recovery tub for recovering cells or tissues held at the interface.
以下、図面を用いて、本発明を実施するための形態(以下、実施形態と称する)を説明する。
Hereinafter, embodiments for carrying out the present invention (hereinafter referred to as embodiments) will be described with reference to the drawings.
図1は、本発明の一実施形態に係わる細胞または組織の輸送装置の外観を示す斜視図である。同図に示す輸送装置1は、アルミニウムやステンレスなどからなる円筒形で大径の外側容器3で全体的に囲まれて構成される。この外側容器3の上端の開口部に、外蓋31が密接に嵌合し、外側容器3内を密閉状態にし得るように構成される。
FIG. 1 is a perspective view showing an external appearance of a cell or tissue transport apparatus according to an embodiment of the present invention. The transport device 1 shown in the figure is configured by being entirely surrounded by a cylindrical and large-diameter outer container 3 made of aluminum, stainless steel, or the like. The outer lid 31 is closely fitted in the opening at the upper end of the outer container 3 so that the inside of the outer container 3 can be sealed.
また、外側容器3の内部に、細胞または組織の保持容器である遠沈管201が配設される。遠沈管201は、後述するように、高比重溶液と低比重溶液との間に形成される界面に細胞を浮遊して保持するように高比重溶液および低比重溶液を収容する。この遠沈管201の下部は、半球状もしくは円錐状に形成される。この遠沈管201の下側にはスプリング33sが配設される。このスプリング33sにより、遠沈管201が緩衝性をもって支持される。
Also, a centrifuge tube 201 which is a cell or tissue holding container is disposed inside the outer container 3. As will be described later, the centrifuge tube 201 accommodates the high specific gravity solution and the low specific gravity solution so as to float and hold the cells at the interface formed between the high specific gravity solution and the low specific gravity solution. The lower part of the centrifuge tube 201 is formed in a hemispherical shape or a conical shape. A spring 33 s is disposed below the centrifuge tube 201. The centrifuge tube 201 is supported by the spring 33s with buffering properties.
更に、外側容器3の中程に、USB35bが電気ケーブルを介して接続される。このUSB35bは、電気ケーブルを介して輸送装置1の内部の後述する制御部に接続されるとともに、図示しない外部の制御装置に接続される。USB35bは、この制御装置からUSB35bを介して、輸送装置1に電源や制御信号を供給するとともに、輸送装置1からの情報を、USB35bを介して外部の制御装置に送信する。また、このUSB35bが接続された部分の少し上方の外側容器3にLEDが設けられる。このLEDは、輸送装置1が作動状態にあるか否かを通知するようにしても良い。
Furthermore, the USB 35b is connected to the middle of the outer container 3 via an electric cable. The USB 35b is connected to a later-described control unit inside the transport apparatus 1 via an electric cable, and is connected to an external control apparatus (not shown). The USB 35b supplies power and control signals from the control device to the transport device 1 via the USB 35b, and transmits information from the transport device 1 to an external control device via the USB 35b. In addition, an LED is provided in the outer container 3 slightly above the portion to which the USB 35b is connected. This LED may notify whether or not the transport device 1 is in an operating state.
図2は、図1に示した輸送装置1の内部構造を示す縦断面を示す模式図である。図2に示すように、遠沈管201の内部に、高比重溶液9と低比重溶液11が入れられる。この遠沈管201内に入れられた2種類の溶液のうち、下方に沈む高比重溶液9とその上方に浮かぶ低比重溶液11との間に、界面13が形成される。この界面13に細胞15が浮遊するようにして保持される。これにより輸送装置1は、細胞15に対して足場が提供されない状態にするとともに、細胞15に対して輸送時の振動などによる物理的衝撃が伝わらず、また細胞の活性にダメージが与えられない構成とされる。
FIG. 2 is a schematic diagram showing a longitudinal section showing the internal structure of the transport apparatus 1 shown in FIG. As shown in FIG. 2, the high specific gravity solution 9 and the low specific gravity solution 11 are placed in the centrifuge tube 201. An interface 13 is formed between the high specific gravity solution 9 that sinks downward and the low specific gravity solution 11 that floats above the two types of solutions placed in the centrifuge tube 201. The cells 15 are held so as to float on the interface 13. As a result, the transport device 1 is in a state in which no scaffold is provided for the cells 15, a physical impact due to vibrations during transport is not transmitted to the cells 15, and the cell activity is not damaged. It is said.
また、回収笊17が、界面13の下方の高比重溶液9内に配設される。また、この回収笊17の周縁部を囲っている笊枠19に、上方から下方に垂直に伸びたチューブ21の下端部が、固定的に取り付けられる。
In addition, the recovery rod 17 is disposed in the high specific gravity solution 9 below the interface 13. Further, the lower end portion of the tube 21 extending vertically downward from above is fixedly attached to the flange frame 19 surrounding the peripheral edge portion of the recovery rod 17.
なお、回収笊17は、例えばメッシュサイズ5μmのナイロンメッシュで構成される。回収笊17は、この回収笊17の上方の界面13に浮遊して保持される細胞15を、後述するように掬い上げて効率良く回収し得るように構成される。また、笊枠19の内側に、タンパク質吸収阻害剤が塗布され、回収笊17への細胞接着が防止される。
Note that the collection basket 17 is made of, for example, a nylon mesh having a mesh size of 5 μm. The collection basket 17 is configured so that the cells 15 suspended and held at the interface 13 above the collection basket 17 can be scooped up and efficiently collected as described later. In addition, a protein absorption inhibitor is applied to the inside of the cocoon frame 19, and cell adhesion to the collection tub 17 is prevented.
遠沈管201の上端部の内側に、内蓋23が嵌合する。この内蓋23は、図3Aおよび図3Bに拡大して示すように、外周面が下方に向かってしぼむ。このしぼんだ下端部は、低比重溶液11内において界面13の上方で界面13に対向する。
The inner lid 23 is fitted inside the upper end of the centrifuge tube 201. As shown in an enlarged view in FIGS. 3A and 3B, the inner lid 23 has an outer peripheral surface that is squeezed downward. The squeezed lower end faces the interface 13 above the interface 13 in the low specific gravity solution 11.
内蓋23は、空気排出部を有する。図3Aおよび図3Bに示すように、空気排出部は、略中央に形成された空気排出孔の下端部から下方に向かって円錐状に広がる。空気排出部は、この広がった下端の開口部が遠沈管201内の空気を収集するように、遠沈管201の内壁に近接する円錐状部23bを有する。この円錐状部23bの周縁部寄りに、遠沈管201内に溶液を注入するための溶液注入孔が形成される。この溶液注入孔にチューブ21の上端が挿入される。
The inner lid 23 has an air discharge part. As shown in FIG. 3A and FIG. 3B, the air discharge portion spreads conically downward from the lower end portion of the air discharge hole formed substantially at the center. The air discharge portion has a conical portion 23 b that is close to the inner wall of the centrifuge tube 201 so that the widened opening at the lower end collects air in the centrifuge tube 201. A solution injection hole for injecting the solution into the centrifuge tube 201 is formed near the peripheral edge of the conical portion 23b. The upper end of the tube 21 is inserted into the solution injection hole.
内蓋23の略中央の空気排出孔には、短い空気排出パイプ23aが挿入される。この空気排出パイプ23aの上端に、封止部23cが形成される。封止部23cは、封止用シーラを使用して空気排出パイプ23aを封止する。また、内蓋23の周縁部寄りを貫通して延出したチューブ21の上端に、封止部21cが形成される。封止部21cは、封止用シーラを使用してチューブ21を封止する。
A short air discharge pipe 23 a is inserted into the air discharge hole at the substantially center of the inner lid 23. A sealing portion 23c is formed at the upper end of the air discharge pipe 23a. The sealing part 23c seals the air discharge pipe 23a using a sealing sealer. Further, a sealing portion 21 c is formed at the upper end of the tube 21 that extends through the periphery of the inner lid 23. The sealing part 21c seals the tube 21 using a sealing sealer.
空気排出パイプ23aおよびチューブ21のそれぞれの上端部は、上述したように、封止用シーラを使用して封止されることにより、内蓋23は、チューブ21から遠沈管201内に溶液を注入しながら空気排出パイプ23aから遠沈管201内の空気を排出し終わった後、空気排出パイプ23aおよびチューブ21から遠沈管201内に空気が入らないように構成される。なお、図示しないが、例えばバルブや密閉蓋などで密閉するように構成されてもよい。封止用シーラを使用して封止したときには、空気排出パイプ23aおよびチューブ21のそれぞれの上端を鋏等によって切断して開封することになる。
As described above, the upper end of each of the air discharge pipe 23a and the tube 21 is sealed using the sealing sealer, so that the inner lid 23 injects the solution from the tube 21 into the centrifuge tube 201. However, after the air in the centrifuge tube 201 is completely discharged from the air discharge pipe 23a, the air is not allowed to enter the centrifuge tube 201 from the air discharge pipe 23a and the tube 21. In addition, although not shown in figure, you may comprise so that it may seal with a valve | bulb, a sealing lid, etc., for example. When the sealing sealer is used for sealing, the upper ends of the air discharge pipe 23a and the tube 21 are cut and opened with scissors or the like.
図2に示すように、遠沈管201の外側に遠沈管201の外周部を全体的に覆うように、フィルムヒータ33g、カーボン33f、アルミテープ33e、アルミ板33dおよびアクリル筒33cが、内側から外側に向かって、記載順に層状に設けられる。また、アクリル筒33cと外側容器3の内周面との間に、断熱と緩衝抑制のために、断熱部33bが設けられる。断熱部33bは、発泡スチロールやアルミニウムビーズ等からなる。なお、遠沈管201の外周部とフィルムヒータ33gとは、図では離間しているが、実際は遠沈管201が挿脱自在な程度に密着する。
As shown in FIG. 2, the film heater 33g, the carbon 33f, the aluminum tape 33e, the aluminum plate 33d, and the acrylic cylinder 33c are arranged from the inside to the outside so as to entirely cover the outer periphery of the centrifuge tube 201 outside the centrifuge tube 201. In the order of description, they are provided in layers. Further, a heat insulating portion 33b is provided between the acrylic cylinder 33c and the inner peripheral surface of the outer container 3 for heat insulation and buffer suppression. The heat insulating portion 33b is made of expanded polystyrene, aluminum beads, or the like. In addition, although the outer peripheral part of the centrifuge tube 201 and the film heater 33g are spaced apart from each other in the drawing, they are in close contact with each other so that the centrifuge tube 201 can be inserted and removed.
更に、外側容器3の上開口部を閉塞している外蓋31の下方であって、空気排出パイプ23aの上側に、温度ロガー31aが設けられる。この温度ロガー31aは、温度を測定し、記録するよう構成される。また、外側容器3の内周面に、例えばワンチップマイクロプロセッサなどからなる制御部35が設けられる。この制御部35に、USB35bが電気ケーブルを介して接続される。アクリル筒33cの外周部に、サーモスタット35aが取り付けられる。このサーモスタット35aは、電気ケーブルを介して制御部35に接続される。
Furthermore, a temperature logger 31a is provided below the outer lid 31 blocking the upper opening of the outer container 3 and above the air discharge pipe 23a. The temperature logger 31a is configured to measure and record the temperature. Further, a control unit 35 made of, for example, a one-chip microprocessor is provided on the inner peripheral surface of the outer container 3. A USB 35b is connected to the control unit 35 via an electric cable. A thermostat 35a is attached to the outer periphery of the acrylic cylinder 33c. The thermostat 35a is connected to the control unit 35 via an electric cable.
以上のように構成される本実施形態の輸送装置1において、CPCに輸送するために医療機関において患者から採取された細胞15は、遠沈管201内に注入された高比重溶液9および低比重溶液11の間に形成された界面13の上に浮遊するように、保持される。
In the transport apparatus 1 of the present embodiment configured as described above, the cells 15 collected from the patient in the medical institution for transport to the CPC are the high specific gravity solution 9 and the low specific gravity solution injected into the centrifuge tube 201. 11 is held so as to float on the interface 13 formed between them.
なお、このように遠沈管201内に、高比重溶液9および低比重溶液11が注入されるとともに、両者の界面13に細胞15が、浮遊して保持される。その後、遠沈管201内の空気を排出するために、チューブ21から高比重溶液9または低比重溶液11が、遠沈管201内に追加注入されながら、遠沈管201内の空気が、円錐状部23bで収集される。この収集された空気が、空気排出パイプ23aから外部に排出され、遠沈管201内の空気が、排出される。それから、遠沈管201の上開口部に内蓋23が気密状態に嵌合されるとともに、チューブ21および空気排出パイプ23aの上端が、封止用シーラで封止される。
In this way, the high specific gravity solution 9 and the low specific gravity solution 11 are injected into the centrifuge tube 201 as described above, and the cells 15 are suspended and held at the interface 13 between them. Thereafter, in order to discharge the air in the centrifuge tube 201, the high specific gravity solution 9 or the low specific gravity solution 11 is additionally injected into the centrifuge tube 201 from the tube 21, and the air in the centrifuge tube 201 is changed into the conical portion 23b. Collected in The collected air is discharged to the outside from the air discharge pipe 23a, and the air in the centrifuge tube 201 is discharged. Then, the inner lid 23 is fitted in an airtight state in the upper opening of the centrifuge tube 201, and the upper ends of the tube 21 and the air discharge pipe 23a are sealed with a sealing sealer.
このようにして高比重溶液9と低比重溶液11との間の界面13に細胞15が、浮遊して保持されるとともに、遠沈管201から空気を抜き、それから内蓋23で遠沈管201内を気密状態にした後、この遠沈管201を外側容器3内に配設する。さらに遠沈管201の下端部が、スプリング33s上に載置される。
In this way, the cells 15 are held in a suspended state at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11, and air is evacuated from the centrifuge tube 201. After the airtight state, the centrifuge tube 201 is disposed in the outer container 3. Further, the lower end portion of the centrifuge tube 201 is placed on the spring 33s.
次に、制御部35は、フィルムヒータ33gを駆動して発熱させる。この発熱により遠沈管201内の高比重溶液9および低比重溶液11を所定の温度、例えば25℃乃至37℃などの温度に加温して、高比重溶液9と低比重溶液11との界面13に保持されている細胞15を当該所定の温度に暖める。それから制御部35は、細胞15を常に所定の温度に保温するように制御し、細胞15の生存率を高い状態を保持する。
Next, the control unit 35 drives the film heater 33g to generate heat. Due to this heat generation, the high specific gravity solution 9 and the low specific gravity solution 11 in the centrifuge tube 201 are heated to a predetermined temperature, for example, a temperature of 25 ° C. to 37 ° C., for example, and the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11. The cell 15 held in is warmed to the predetermined temperature. Then, the control unit 35 controls to keep the cells 15 at a predetermined temperature at all times, and keeps the survival rate of the cells 15 high.
上述したように、本実施形態の輸送装置1は、遠沈管201内で界面13に細胞15を浮遊して保持するとともに、細胞15を所定の温度に保温しつつ、例えば宅配便などによりCPCに輸送される。本実施形態の輸送装置1は、高比重溶液9と低比重溶液11を入れた遠沈管201を外側容器3内に収納するという極めてコンパクトで簡便な構造であって、輸送における取扱いが極めて簡単なものである。この輸送では、空気が排出された気密状態の遠沈管201内において高比重溶液9と低比重溶液11との間の界面13に細胞15が浮遊して保持されるため、輸送時の細胞への振動などの衝撃および細胞活性へのダメージを軽減することができる。さらに本実施形態の輸送装置1は、足場を提供しないため、細胞の増殖活性を停止または低下することにより、細胞増殖による細胞保持容器内の環境悪化の防止も可能であると同時に、細胞を休眠状態に保持することができる。
As described above, the transport device 1 according to the present embodiment floats and holds the cells 15 on the interface 13 in the centrifuge tube 201 and keeps the cells 15 at a predetermined temperature, for example, by courier or the like. Transported. The transport device 1 of this embodiment has an extremely compact and simple structure in which the centrifuge tube 201 containing the high specific gravity solution 9 and the low specific gravity solution 11 is housed in the outer container 3, and is extremely easy to handle in transportation. Is. In this transportation, since the cells 15 are suspended and held at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 in the airtight centrifuge tube 201 from which the air has been exhausted, Impacts such as vibration and damage to cell activity can be reduced. Furthermore, since the transport device 1 of the present embodiment does not provide a scaffold, it is possible to prevent environmental deterioration in the cell holding container due to cell proliferation by stopping or reducing cell proliferation activity, and at the same time, dormant cells. Can be kept in a state.
また、細胞15は、制御部35の制御による加温制御により常に所定の最適温度に保持されているため、従来のような低温傷害が生ぜず、細胞活性を維持した状態で輸送されることができる。
In addition, since the cells 15 are always maintained at a predetermined optimum temperature by the heating control under the control of the control unit 35, the cells 15 can be transported in a state in which the cell activity is maintained without causing a low-temperature injury as in the prior art. it can.
更に、輸送装置1をCPCに輸送した後、CPCにおいて細胞15から組織を生産するために、細胞輸送装置1から細胞15を回収することが必要となる。この細胞15の回収において、細胞輸送装置1の外側容器3の外蓋31が取り外されて、外側容器3から遠沈管201が取り出される。この取り出された遠沈管201から、内蓋23が上方に持ち上げるように取り出されると、この内蓋23に固定されているチューブ21も同時に上方に持ち上げられる。このチューブ21の下端部に固定されている回収笊17も高比重溶液9および低比重溶液11をその網目から排液しながら同時に上方に持ち上げられて取り出される。従って、高比重溶液9と低比重溶液11との界面13に保持された細胞15は、回収笊17で掬い上げられるように簡単に回収されることになる。
Furthermore, after the transport device 1 is transported to the CPC, it is necessary to collect the cells 15 from the cell transport device 1 in order to produce tissue from the cells 15 in the CPC. In the collection of the cells 15, the outer lid 31 of the outer container 3 of the cell transport device 1 is removed, and the centrifuge tube 201 is taken out from the outer container 3. When the inner lid 23 is removed from the removed centrifuge tube 201 so as to be lifted upward, the tube 21 fixed to the inner lid 23 is also lifted upward. The recovery rod 17 fixed to the lower end portion of the tube 21 is also lifted and taken out simultaneously while draining the high specific gravity solution 9 and the low specific gravity solution 11 from the mesh. Therefore, the cells 15 held at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 are easily recovered so as to be scooped up by the recovery rod 17.
このように細胞15は、単に遠沈管201から内蓋23を上方に持ち上げることにより、非常に簡単に回収されることができる。また、本実施形態の輸送装置1では、高比重溶液9にパーフルオロカーボン(PFC)を用いるため、低比重溶液のガス制御、すなわちPFCと培地または保存溶液との間のガス移動による制御を行うことができる。これにより輸送装置1は、温度およびpHなどの通常の培養条件を用いることで輸送時の細胞活性へのダメージを軽減することを可能とする。
Thus, the cells 15 can be recovered very simply by simply lifting the inner lid 23 upward from the centrifuge tube 201. Further, in the transport device 1 of the present embodiment, since perfluorocarbon (PFC) is used for the high specific gravity solution 9, gas control of the low specific gravity solution, that is, control by gas movement between the PFC and the culture medium or the storage solution is performed. Can do. Thereby, the transport apparatus 1 makes it possible to reduce damage to cell activity during transport by using normal culture conditions such as temperature and pH.
図4および図5は、本発明の他の実施形態に係わる細胞または組織の輸送装置に使用される遠沈管201を示す斜視図である。図5は、下部の遠沈管201とその上部とが切り離されて分離した状態を図示する。図4および図5に示す遠沈管201は、その内部に高比重溶液9と低比重溶液11とが入れられる。この2種類の溶液のうち、下方に沈む高比重溶液9とその上方に浮かぶ低比重溶液11との間に、界面13が図4に示すように形成される。この界面13に細胞15が浮遊して保持され、細胞15に対して足場がない状態を形成する。これにより、細胞15に対して輸送時の振動などによる物理的衝撃が伝達せず、また細胞活性にダメージが与えられないように構成される。
4 and 5 are perspective views showing a centrifuge tube 201 used in a cell or tissue transport apparatus according to another embodiment of the present invention. FIG. 5 illustrates a state where the lower centrifuge tube 201 is separated from the upper portion thereof. The centrifuge tube 201 shown in FIG. 4 and FIG. 5 contains the high specific gravity solution 9 and the low specific gravity solution 11 therein. Of these two types of solutions, an interface 13 is formed as shown in FIG. 4 between the high specific gravity solution 9 sinking downward and the low specific gravity solution 11 floating above. The cells 15 are suspended and held on the interface 13 and form a state where there is no scaffold for the cells 15. Thereby, the physical impact by the vibration at the time of transport etc. is not transmitted with respect to the cell 15, and it is comprised so that a cell activity may not be damaged.
なお、図5および図9に示すように、遠沈管201の上端部外側に、ねじ部201aが形成される。このねじ部201aに、遠沈管201の上開口部を緊密に閉塞している内蓋213と接着剤等により固定的に取り付けられ固着される内蓋キャップ123dの内側に形成されたねじ部123eが螺合する。図6は、この螺合した状態を図示する。
In addition, as shown in FIGS. 5 and 9, a screw portion 201 a is formed outside the upper end portion of the centrifuge tube 201. A screw portion 123e formed on the inner side of the inner lid cap 123d fixedly attached and fixed to the inner lid 213 tightly closing the upper opening of the centrifuge tube 201 with an adhesive or the like is attached to the screw portion 201a. Screw together. FIG. 6 illustrates this screwed state.
図7Aおよび図7Bに示すように、内蓋213の中央に形成された円筒部225aの下端部から下方に向かって、円錐状広がり部225が、円錐状に広がるように形成される。この広がった下端の開口部が、遠沈管201内の空気を収集する。
As shown in FIGS. 7A and 7B, a conical spreading portion 225 is formed so as to spread in a conical shape downward from a lower end portion of a cylindrical portion 225a formed at the center of the inner lid 213. This widened opening at the lower end collects air in the centrifuge tube 201.
更に、内蓋213と、この内蓋213に固着される内蓋キャップ123dの中央部に、空気抜きパイプ123aが貫通して取り付けられる。この空気抜きパイプ123aの上端部に、空気抜き部用キャップ123cが緊密に嵌合されることにより、空気抜きパイプ123aが緊密に閉塞される。なお、この内蓋キャップ123dが嵌合する空気抜きパイプ123aの上部は、図2に示すように、空気抜き部用キャップ123cの代わりにシリンジによって閉塞されるように構成されても良い。
Furthermore, an air vent pipe 123a is attached through the inner lid 213 and the central portion of the inner lid cap 123d fixed to the inner lid 213. The air vent pipe 123a is tightly closed by tightly fitting the air vent cap 123c to the upper end of the air vent pipe 123a. In addition, as shown in FIG. 2, the upper part of the air vent pipe 123a to which this inner lid cap 123d is fitted may be configured to be closed by a syringe instead of the air vent cap 123c.
空気抜きパイプ123aの下端部は、内蓋213内の円錐状広がり部225の上部の円筒部225aの上端部に連結される。両者内を、すなわち空気抜きパイプ123aと円筒部225a内を、空気や溶液が、通過し得るように構成される。
The lower end portion of the air vent pipe 123a is connected to the upper end portion of the cylindrical portion 225a above the conical spreading portion 225 in the inner lid 213. Air or a solution can pass through both of them, that is, through the air vent pipe 123a and the cylindrical portion 225a.
内蓋キャップ123dの周縁部寄りに、チューブ121の上端が貫通して設けられる。このチューブ121の上端部に、チューブ用キャップ121cが、緊密に嵌合するように取り付けられる。チューブ121の下部は、内蓋キャップ123dを貫通し、さらに内蓋213内の後述する回収笊117の外側を、すなわち内蓋213内の円錐状広がり部225の周縁部寄りに図7Aおよび図7Bに示すように形成されたチューブ孔123fを通って下方に延出する。チューブ121の下端部は、遠沈管201内に下方まで伸びている。そして、チューブ用キャップ121cを取り外した場合に、チューブ121の上端から取り入れられる外部の空気がチューブ121を通って、遠沈管201内に取り込まれ得るように構成される。
The upper end of the tube 121 is provided so as to penetrate near the peripheral edge of the inner lid cap 123d. A tube cap 121c is attached to the upper end portion of the tube 121 so as to be tightly fitted. The lower part of the tube 121 penetrates the inner lid cap 123d, and further outside the recovery rod 117 described later in the inner lid 213, that is, near the peripheral edge of the conical spreading portion 225 in the inner lid 213. It extends downward through the tube hole 123f formed as shown in FIG. The lower end of the tube 121 extends downward into the centrifuge tube 201. When the tube cap 121c is removed, external air taken in from the upper end of the tube 121 can be taken into the centrifuge tube 201 through the tube 121.
内蓋213の円錐状広がり部225の途中に、図7Aおよび図7Bにも示すように、回収笊117が配設される。この回収笊117は、円錐状広がり部225の傾斜部に上側角部が食い込むように配設される。また、回収笊117の周縁部に、図8Aおよび図8Bに示すように、笊枠119が設けられる。なお、回収笊117の網目状部は、例えばメッシュサイズ5μmのナイロンメッシュで構成される。回収笊117の網目状部は、この回収笊117で高比重溶液9と低比重溶液11の界面13に浮遊して保持されている細胞15を回収し得るように構成される。また、笊枠119および遠沈管201の内側に、タンパク質吸収阻害剤が塗布され、これにより回収笊117への細胞接着を防止する。
As shown in FIG. 7A and FIG. 7B, a collection basket 117 is disposed in the middle of the conical spreading portion 225 of the inner lid 213. The collection basket 117 is arranged such that the upper corner bites into the inclined part of the conical spreading part 225. Further, as shown in FIG. 8A and FIG. 8B, a collar frame 119 is provided on the peripheral edge of the collection basket 117. The mesh portion of the collection basket 117 is made of, for example, a nylon mesh having a mesh size of 5 μm. The mesh portion of the collection basket 117 is configured so that the collection basket 117 can collect the cells 15 suspended and held at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11. In addition, a protein absorption inhibitor is applied to the inside of the gutter frame 119 and the centrifuge tube 201, thereby preventing cell adhesion to the collection gutter 117.
以上のように構成される本実施形態の遠沈管201は、当該遠沈管201内の高比重溶液9と低比重溶液11との界面13に細胞15を浮遊させながら保持する。さらに、内部の溶液が漏出しないように、内蓋キャップ123dを緊密に嵌合した後、遠沈管201は、輸送装置1内に収納され、CPCに輸送される。
The centrifuge tube 201 of the present embodiment configured as described above holds the cells 15 while floating at the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 in the centrifuge tube 201. Furthermore, after the inner lid cap 123d is tightly fitted so that the solution inside does not leak, the centrifuge tube 201 is housed in the transport device 1 and transported to the CPC.
この遠沈管201を収納した輸送装置1を受け取ったCPCは、細胞15から組織を生産するために、細胞15を遠沈管201から回収する。この回収において、まず空気抜き部用キャップ123cおよびチューブ用キャップ121cが取り外され、空気抜き部用キャップ123cの代わり空気抜きパイプ123aの上端に図示しないシリンジが取り付けられる。
The CPC that has received the transport device 1 containing the centrifuge tube 201 collects the cell 15 from the centrifuge tube 201 in order to produce tissue from the cell 15. In this recovery, the air vent cap 123c and the tube cap 121c are first removed, and a syringe (not shown) is attached to the upper end of the air vent pipe 123a instead of the air vent cap 123c.
それから、遠沈管201を逆さまにし、空気抜きパイプ123aに取り付けたシリンジで高比重溶液9および低比重溶液11が抜き取られる。遠沈管201内の高比重溶液9と低比重溶液11が抜き取られると、これらの溶液の界面13に浮遊して保持されていた細胞15は、回収笊117の上に乗るように回収される。
Then, the centrifuge tube 201 is turned upside down, and the high specific gravity solution 9 and the low specific gravity solution 11 are extracted with a syringe attached to the air vent pipe 123a. When the high specific gravity solution 9 and the low specific gravity solution 11 in the centrifuge tube 201 are extracted, the cells 15 suspended and held at the interface 13 of these solutions are collected so as to ride on the collection basket 117.
このように回収笊117が細胞15を回収した後、新しい遠沈管201が取り付けられ、逆さまの状態から元の正常な状態に戻し、空気抜きパイプ123aに取り付けたシリンジから、リン酸緩衝生理食塩水(PBS)が遠沈管201内に流し込まれる。これにより回収笊117に回収された細胞15は、遠沈管201内に流し入れられる。細胞15が新たな遠沈管201内に流し込まれ、遠沈管201に新しい蓋がされた後、遠沈管201が遠心分離機にかけられ、細胞15が回収される。
After the recovery rod 117 collects the cells 15 in this manner, a new centrifuge tube 201 is attached, and the normal state is restored from the upside-down state. From the syringe attached to the air vent pipe 123a, phosphate buffered saline ( PBS) is poured into the centrifuge tube 201. As a result, the cells 15 collected in the collection basket 117 are poured into the centrifuge tube 201. After the cells 15 are poured into a new centrifuge tube 201 and the centrifuge tube 201 is covered with a new lid, the centrifuge tube 201 is centrifuged and the cells 15 are collected.
図10は、本発明の別の実施形態に係わる細胞または組織の輸送装置に使用される細胞収納容器の縦断面を示す模式図である。同図に示す細胞収納容器200は、容器本体222の上開口部と上蓋223dの内側に形成されたねじとが螺合して、容器本体222を密閉状態で閉塞し得るように構成される。
FIG. 10 is a schematic view showing a longitudinal section of a cell storage container used in a cell or tissue transport apparatus according to another embodiment of the present invention. The cell storage container 200 shown in the figure is configured such that the upper opening of the container main body 222 and a screw formed inside the upper lid 223d are screwed together to close the container main body 222 in a sealed state.
容器本体222の外側に、フィルムヒータ233fが設けられ、フィルムヒータ233fの外側に断熱層233bが設けられる。この構成により容器本体222内を所定の温度に加温し得るように構成される。なお、フィルムヒータ233fは、密着して設けられ容器と一体的に構成される。
A film heater 233f is provided outside the container body 222, and a heat insulating layer 233b is provided outside the film heater 233f. With this configuration, the inside of the container main body 222 can be heated to a predetermined temperature. In addition, the film heater 233f is provided in close contact and is configured integrally with the container.
上蓋223dの下面に、上端が近接して円筒部311が設けられる。この円筒部311の下部は、容器本体222の内蓋223として形成される。また、内蓋223は、円筒部311の下端部から下方に向かって円錐状に広がった円錐状広がり部223bを形成する。この円錐状広がり部223bの下方の円錐状に広がった下端の開口部は、容器本体222の内周壁に内溶液が漏れない程度にかつ着脱自在であるように密接する。
A cylindrical portion 311 is provided on the lower surface of the upper lid 223d with its upper end close to it. A lower portion of the cylindrical portion 311 is formed as an inner lid 223 of the container main body 222. Further, the inner lid 223 forms a conical spreading portion 223b that spreads conically downward from the lower end portion of the cylindrical portion 311. The opening at the lower end that expands in a conical shape below the conical expanding portion 223b is in close contact with the inner peripheral wall of the container body 222 so that the inner solution does not leak and is detachable.
内蓋223の中央部には、空気抜きパイプ311が貫通して取り付けられる。この空気抜きパイプ311の上端部に、空気抜き部用キャップ233cが緊密に嵌合され、空気抜きパイプ311を緊密に閉塞する。なお、この空気抜きパイプ311の上部は、空気抜き部用キャップ233cの代わりに図2に示す封止用シーラを使用して封止または閉塞するようにしても良い。
An air vent pipe 311 is attached through the central portion of the inner lid 223. An air vent cap 233c is tightly fitted to the upper end of the air vent pipe 311 to close the air vent pipe 311 tightly. The upper part of the air vent pipe 311 may be sealed or closed using a sealing sealer shown in FIG. 2 instead of the air vent cap 233c.
円錐状広がり部223bの下方の容器本体222の内部に、高比重溶液9と低比重溶液11が入れられる。この容器本体222内に入れられた2種類の溶液のうち、下方に沈む高比重溶液9とその上方に浮かぶ低比重溶液11との間には界面13が形成される。この界面13に培養された皮膚組織等の細胞シートである組織片215が浮遊して保持される。組織片215に対して足場がない状態が形成されるとともに、組織片215に対して輸送時の振動などによる物理的衝撃が伝達せず、また細胞活性にダメージが与えられないように構成される。
The high specific gravity solution 9 and the low specific gravity solution 11 are placed in the container main body 222 below the conical spreading portion 223b. An interface 13 is formed between the high specific gravity solution 9 that sinks downward and the low specific gravity solution 11 that floats above the two types of solutions placed in the container body 222. A tissue piece 215 which is a cell sheet such as a skin tissue cultured on the interface 13 is suspended and held. The tissue piece 215 is configured such that no scaffold is formed, the physical shock due to vibration during transportation is not transmitted to the tissue piece 215, and the cell activity is not damaged. .
上述したように構成される本実施形態の細胞収納容器200において、容器本体222が全体的に内蓋223で閉塞される。また上蓋223dで上開口部が密閉状態で閉塞された容器本体222内に、高比重溶液9と低比重溶液11が入れられ、この高比重溶液9と低比重溶液11との間の界面13に組織片215が、浮遊して保持される。これにより、この細胞収納容器200の輸送時にも振動などによる物理的衝撃が組織片215に伝達することなく、安全に組織片215が輸送されることができる。
In the cell storage container 200 of the present embodiment configured as described above, the container body 222 is entirely closed with the inner lid 223. Further, the high specific gravity solution 9 and the low specific gravity solution 11 are placed in the container main body 222 whose upper opening is closed in a sealed state by the upper lid 223d, and the interface 13 between the high specific gravity solution 9 and the low specific gravity solution 11 is placed in the container body 222. The tissue piece 215 is held floating. Thereby, the tissue piece 215 can be safely transported without transferring a physical impact due to vibration or the like to the tissue piece 215 even when the cell storage container 200 is transported.
上述したように、本実施形態の細胞または組織の輸送装置1は、細胞または組織を搬送する際に用いられる細胞および組織の輸送装置である。輸送装置1は、界面が形成される高比重溶液と低比重溶液とを収容する遠沈管201(保持容器)と、界面に保持される細胞または組織とは離隔して遠沈管201(保持容器)に配設され、界面に保持された細胞または組織を回収するための回収笊17とを有する。
As described above, the cell or tissue transport apparatus 1 of this embodiment is a cell and tissue transport apparatus used when transporting cells or tissues. The transport apparatus 1 includes a centrifuge tube 201 (holding container) that accommodates a high density solution and a low specific gravity solution in which an interface is formed, and a cell or tissue held at the interface, and the centrifuge tube 201 (holding container). And a recovery rod 17 for recovering cells or tissue held at the interface.
このような輸送装置1は、高比重溶液と低比重溶液との間に形成される界面に細胞または組織を保持するとともに、界面に保持される細胞から離隔して回収笊17を配設し、この回収笊17で界面に保持された細胞を回収する。これにより、本実施形態の輸送装置1は、輸送時の振動などによる物理的衝撃が細胞に伝達されず、細胞活性にダメージが与えられることもなく、細胞または組織を安全に輸送できる。さらに本実施形態の輸送装置1は、界面に保持されている細胞または組織を簡単かつ効率良く回収することができる。
Such a transport device 1 holds cells or tissues at the interface formed between the high specific gravity solution and the low specific gravity solution, and disposes a collection basket 17 apart from the cells held at the interface, The cells held at the interface are recovered by the recovery rod 17. As a result, the transport device 1 of the present embodiment can safely transport cells or tissues without causing physical impact due to vibration during transport to the cells and without damaging the cell activity. Furthermore, the transport device 1 of the present embodiment can easily and efficiently collect cells or tissues held at the interface.
また輸送装置1はさらに、遠沈管201(保持容器)を密閉すべく、遠沈管201(保持容器)の上開口部を閉鎖する内蓋23(容器蓋)と、空気排出部と、空気排出パイプ23aと、チューブ21(溶液注入パイプ)とを有する。
Further, the transport apparatus 1 further includes an inner lid 23 (container lid) for closing the upper opening of the centrifuge tube 201 (holding container), an air discharge unit, and an air discharge pipe so as to seal the centrifuge tube 201 (holding container). 23a and a tube 21 (solution injection pipe).
空気排出部は、空気排出孔、円錐状部23b(円錐状広がり部)および溶液注入孔を有する。空気排出孔は、この内蓋23(容器蓋)の下側に配設され、略中央に形成される。円錐状部23b(円錐状広がり部)は、この空気排出孔の下端部から下方に向かって円錐状に広がり、この広がった下端の開口部が遠沈管201(保持容器)の空気を収集するように、遠沈管201(保持容器)の内壁に近接して配設される。溶液注入孔は、この円錐状部23b(円錐状広がり部)の周縁部寄りに形成され、遠沈管201(保持容器)内に溶液を投入するために配設される。
The air discharge part has an air discharge hole, a conical part 23b (conical spreading part) and a solution injection hole. The air discharge hole is disposed below the inner lid 23 (container lid) and is formed substantially at the center. The conical portion 23b (conical spreading portion) spreads conically downward from the lower end of the air discharge hole, and the opening at the lower end of the air collecting collects the air in the centrifuge tube 201 (holding container). The centrifuge tube 201 (holding container) is disposed close to the inner wall. The solution injection hole is formed near the peripheral edge of the conical portion 23b (conical spreading portion), and is arranged for introducing the solution into the centrifuge tube 201 (holding container).
空気排出パイプ23aは、空気排出孔内に下端が挿通され、上端が上方に突出して容器蓋の下面の略中央に形成された円筒状中央凹部に密接に嵌合する。チューブ21(溶液注入パイプ)は、上端が溶液注入孔を貫通して上方に突出し内蓋23(容器蓋)の下面の周縁寄りに形成された円筒状周縁凹部に密接に嵌合し、下端が下方に延出し、回収笊17を固定的に保持する。
The lower end of the air discharge pipe 23a is inserted into the air discharge hole, the upper end protrudes upward, and is closely fitted into a cylindrical central recess formed at the approximate center of the lower surface of the container lid. The tube 21 (solution injection pipe) has an upper end that protrudes upward through the solution injection hole and closely fits into a cylindrical peripheral recess formed near the periphery of the lower surface of the inner lid 23 (container lid). It extends downward and holds the collection basket 17 fixedly.
本実施形態の輸送装置1は、遠沈管201を閉鎖する内蓋23の下側に空気排出部を配設し、この空気排出部の中央の空気排出孔から下方に向かって円錐状に広がる円錐状部23bを設ける。本実施形態の輸送装置1は、この円錐状部23bの周縁部寄りの溶液注入孔に通したチューブ21から遠沈管201内に溶液を注入しながら、遠沈管201内の空気を円錐状部23bで収集し、空気排出孔に通した空気排出パイプ23aから空気を排出する。溶液注入孔に通したチューブ21の下端に回収笊17を固定的に保持しているため、遠沈管201内の空気は、排出されることができる。これにより本実施形態の輸送装置1は、輸送時の振動などによる遠沈管201内の溶液の揺れを低減し、細胞または組織への物理的衝撃を軽減できる。また本実施形態の輸送装置1は、チューブ21の下端に回収笊17が固定的に保持されているため、空気排出部をチューブ21とともに持ち上げることにより回収笊17を容易に取り出し、細胞を容易に回収することができる。
In the transport device 1 of the present embodiment, an air discharge portion is disposed below the inner lid 23 that closes the centrifuge tube 201, and a cone that extends downward in a conical shape from the air discharge hole at the center of the air discharge portion. A shaped portion 23b is provided. The transport device 1 of the present embodiment allows the air in the centrifuge tube 201 to flow into the centrifuge portion 23b while injecting the solution into the centrifuge tube 201 from the tube 21 that has passed through the solution injection hole near the periphery of the conical portion 23b. Then, air is discharged from the air discharge pipe 23a collected through the air discharge hole. Since the recovery rod 17 is fixedly held at the lower end of the tube 21 passed through the solution injection hole, the air in the centrifuge tube 201 can be discharged. Thereby, the transport apparatus 1 according to the present embodiment can reduce the shaking of the solution in the centrifuge tube 201 due to vibration during transportation, and can reduce physical impact on cells or tissues. Further, in the transport device 1 of the present embodiment, since the collection basket 17 is fixedly held at the lower end of the tube 21, the collection basket 17 can be easily taken out by lifting the air discharge part together with the tube 21, and the cells can be easily removed. It can be recovered.
本実施形態の輸送装置1は、遠沈管201(保持容器)の周壁部に形成され、該遠沈管201(保持容器)内の温度を所定の温度に加温するためのフィルムヒータ33g(ヒータ)を有する。
The transport apparatus 1 of the present embodiment is formed on a peripheral wall portion of a centrifuge tube 201 (holding container), and a film heater 33g (heater) for heating the temperature in the centrifuge tube 201 (holding container) to a predetermined temperature. Have
本実施形態の輸送装置1は、遠沈管201の周壁部にフィルムヒータ33gを形成し、このフィルムヒータ33gで遠沈管201の温度を所定の温度に加温する。これにより遠沈管201の内部は所定の温度に加温され、高比重溶液と低比重溶液との界面に保持されている細胞または組織も所定の温度に加温され、細胞の生存率を高い状態に保持している。
The transport device 1 of the present embodiment forms a film heater 33g on the peripheral wall portion of the centrifuge tube 201, and heats the temperature of the centrifuge tube 201 to a predetermined temperature with the film heater 33g. As a result, the inside of the centrifuge tube 201 is heated to a predetermined temperature, and cells or tissues held at the interface between the high specific gravity solution and the low specific gravity solution are also heated to the predetermined temperature, and the cell survival rate is high. Hold on.
本実施形態の輸送装置1は、高比重溶液が、パーフルオロカーボンまたは変性シリコーンオイルであり、低比重溶液は、細胞培養用の培地または生理的食塩水である。
In the transport device 1 of this embodiment, the high specific gravity solution is perfluorocarbon or modified silicone oil, and the low specific gravity solution is a cell culture medium or physiological saline.
本実施形態の輸送装置1は、パーフルオロカーボンまたは変性シリコーンオイルの高比重溶液と細胞培養用の培地または生理的食塩水の低比重溶液とを遠沈管201に入れ、高比重溶液と低比重溶液との間の界面に細胞を浮遊させて保持している。これにより本実施形態の輸送装置1は、輸送時の細胞または組織への振動などの物理的衝撃および細胞活性へのダメージを軽減することができる。さらに本実施形態の輸送装置1は、足場を提供しないため、細胞の増殖活性を停止または低下することにより、細胞増殖による遠沈管201内の環境悪化の防止も可能とする。
The transport apparatus 1 according to the present embodiment includes a high specific gravity solution of perfluorocarbon or modified silicone oil and a low specific gravity solution of a cell culture medium or physiological saline in a centrifuge tube 201, and a high specific gravity solution and a low specific gravity solution Cells are suspended and held at the interface between. Thereby, the transport apparatus 1 of the present embodiment can reduce physical impacts such as vibrations to cells or tissues during transport and damage to cell activity. Furthermore, since the transport apparatus 1 of this embodiment does not provide a scaffold, it is possible to prevent deterioration of the environment in the centrifuge tube 201 due to cell proliferation by stopping or reducing cell proliferation activity.
本実施形態の輸送装置1は、回収笊17が、ナイロンメッシュからなる網目状部分とこの網目状部分の外周縁部を囲む笊枠119で構成されていることを要旨とする。
The gist of the transport device 1 of the present embodiment is that the collection basket 17 is configured by a mesh portion made of nylon mesh and a collar frame 119 surrounding the outer peripheral edge of the mesh portion.
本実施形態の輸送装置1は、回収笊17の網目状部分がナイロンメッシュで構成され、回収笊17の外周縁部が笊枠119で囲まれているため、高比重溶液と低比重溶液を除去し、細胞または組織を効率良く回収することができる。
In the transport device 1 of the present embodiment, the mesh portion of the collection basket 17 is made of nylon mesh, and the outer peripheral edge of the collection basket 17 is surrounded by the fence frame 119, so that the high specific gravity solution and the low specific gravity solution are removed. In addition, cells or tissues can be recovered efficiently.
本実施形態の輸送装置1は、笊枠119の内壁にタンパク質吸着阻害剤を塗布する。
The transport device 1 according to this embodiment applies a protein adsorption inhibitor to the inner wall of the frame 119.
本実施形態の輸送装置1は、回収笊17を囲む笊枠19の内壁にタンパク質吸着阻害剤を塗布するため、回収笊17への細胞接着を防止することができる。
Since the transport apparatus 1 of the present embodiment applies the protein adsorption inhibitor to the inner wall of the cage frame 19 that surrounds the recovery basket 17, cell adhesion to the recovery basket 17 can be prevented.
本実施形態の輸送装置1は、高比重溶液と低比重溶液との間に形成される界面に細胞を浮遊して保持するとともに、界面から離隔して回収笊を配設し、この回収笊で界面に保持された細胞を回収する。これにより、本実施形態の輸送装置1は、輸送時の振動などによる物理的衝撃が細胞に伝達されず、細胞活性にダメージが与えられることもなく、細胞を安全に輸送できるとともに、界面に浮遊して保持されている細胞を簡単かつ効率良く回収することができる。
The transport device 1 of the present embodiment floats and holds cells at the interface formed between the high specific gravity solution and the low specific gravity solution, and disposes a recovery basket away from the interface. Collect the cells retained at the interface. As a result, the transport device 1 according to the present embodiment is capable of transporting cells safely without causing physical impact due to vibration during transport to the cells, damaging cell activity, and floating at the interface. Thus, the retained cells can be collected easily and efficiently.
また、本実施形態の輸送装置1は、細胞保持容器を閉鎖する容器蓋の下側に空気排出部を配設し、この空気排出部の中央の空気排出孔から下方に向かって円錐状に広がる円錐状広がり部を設ける。本実施形態の輸送装置1は、この円錐状広がり部の周縁部寄りの溶液注入孔に通した溶液注入パイプから細胞保持容器内に溶液を注入しながら、容器内の空気を円錐状広がり部で収集し、空気排出孔に通した空気排出パイプから空気を排出するとともに、溶液注入孔に通した溶液注入パイプの下端に回収笊を固定的に保持している。これにより本実施形態の輸送装置1は、細胞保持容器内の空気を排出することができ、輸送時の振動などによる細胞保持容器内の溶液の揺れが低減し、細胞への物理的衝撃を軽減できる。さらに本実施形態の輸送装置1は、溶液注入パイプの下端に回収笊が固定的に保持されているので、空気排出部を溶液注入パイプとともに持ち上げることにより回収笊を容易に取り出し、細胞を容易に回収することができる。
Moreover, the transport apparatus 1 of this embodiment arrange | positions an air discharge part below the container lid which closes a cell holding container, and spreads conically downward from the air discharge hole of the center of this air discharge part. A conical expanse is provided. The transport device 1 of the present embodiment injects the air in the container at the conical spreading part while injecting the solution into the cell holding container from the solution injection pipe passing through the solution injection hole near the peripheral edge of the conical spreading part. The collected air is discharged from the air discharge pipe that is passed through the air discharge hole, and the recovery tank is fixedly held at the lower end of the solution injection pipe that is passed through the solution injection hole. As a result, the transport device 1 of the present embodiment can discharge the air in the cell holding container, reduce the shaking of the solution in the cell holding container due to vibration during transportation, and reduce the physical impact on the cell. it can. Further, in the transport device 1 of the present embodiment, since the recovery tub is fixedly held at the lower end of the solution injection pipe, the recovery tub can be easily taken out by lifting the air discharge portion together with the solution injection pipe, and the cells can be easily removed. It can be recovered.
更に、本実施形態の輸送装置1は、細胞保持容器の周壁部にヒータを形成し、このヒータで細胞保持容器内の温度を所定の温度に加温する。これにより本実施形態の輸送装置1は、細胞保持容器の内部は所定の温度に加温され、高比重溶液と低比重溶液との界面に保持されている細胞も所定の温度に加温され、細胞の生存率を高い状態に保持することができる。
Furthermore, the transport device 1 of the present embodiment forms a heater on the peripheral wall portion of the cell holding container, and heats the temperature in the cell holding container to a predetermined temperature with this heater. Thereby, in the transport device 1 of the present embodiment, the inside of the cell holding container is heated to a predetermined temperature, and the cells held at the interface between the high specific gravity solution and the low specific gravity solution are also heated to the predetermined temperature, Cell viability can be kept high.
本実施形態の輸送装置1は、パーフルオロカーボンまたは変性シリコーンオイルなどの高比重溶液と細胞培養用の培地または細胞保存液(例えば生理的食塩水)などの低比重溶液とを細胞保持容器に入れ、高比重溶液と低比重溶液との間の界面に細胞を浮遊させて保持している。これにより本実施形態の輸送装置1は、輸送時の細胞への振動などの物理的衝撃および細胞活性へのダメージを軽減することができる。さらに本実施形態の輸送装置1は、足場を提供しないので、細胞の増殖活性を停止または低下することにより、細胞増殖による細胞保持容器内の環境悪化の防止も可能である。
The transport device 1 of the present embodiment puts a high specific gravity solution such as perfluorocarbon or modified silicone oil and a low specific gravity solution such as a cell culture medium or cell preservation solution (for example, physiological saline) in a cell holding container, Cells are suspended and held at the interface between the high density solution and the low density solution. Thereby, the transport apparatus 1 of this embodiment can reduce physical impacts such as vibrations to cells during transport and damage to cell activity. Furthermore, since the transport device 1 of the present embodiment does not provide a scaffold, it is possible to prevent deterioration of the environment in the cell holding container due to cell growth by stopping or reducing the cell growth activity.
また、本実施形態の輸送装置1は、回収笊の網目状の部分がナイロンメッシュで構成され、回収笊の外周縁部を笊枠で囲まれているので、高比重溶液と低比重溶液を除去し、細胞を効率良く回収することができる。
In addition, the transport device 1 of the present embodiment removes the high specific gravity solution and the low specific gravity solution because the net-like portion of the recovery basket is made of nylon mesh and the outer peripheral edge of the recovery basket is surrounded by a fence frame. Thus, the cells can be collected efficiently.
更に、本実施形態の輸送装置1は、回収笊を囲む笊枠の内壁にタンパク質吸着阻害剤を塗布するので、回収笊への細胞接着を防止することができる。
Furthermore, since the transport apparatus 1 of the present embodiment applies the protein adsorption inhibitor to the inner wall of the cage frame surrounding the recovery basket, cell adhesion to the recovery basket can be prevented.
以上、実施形態を挙げて本発明の実施形態を説明したが、実施形態は例示であり、請求の範囲に記載される発明の範囲は、発明の要旨を逸脱しない範囲内で種々変更できるものである。
The embodiments of the present invention have been described with reference to the embodiments. However, the embodiments are exemplifications, and the scope of the invention described in the claims can be variously changed without departing from the scope of the invention. is there.
1 輸送装置
3 外側容器
9 高比重溶液
11 低比重溶液
13 界面
15 細胞
17 回収笊
19 笊枠
21 チューブ
21c 封止部
23 内蓋
23a 空気排出パイプ
23b 円錐状部(円錐状広がり部)
23c 封止部
31 外蓋
33b 断熱部
33c アクリル筒
33d アルミ板
33e アルミテープ
33f カーボン
33g フィルムヒータ
33s スプリング
35 制御部
35b USB
117 回収笊
121 チューブ(溶液注入パイプ)
121c チューブ用キャップ
123a 空気抜きパイプ
123c 空気抜き部用キャップ
123d 内蓋キャップ
200 細胞収納容器
201 遠沈管(保持容器)
213 内蓋(容器蓋)
215 組織片
222 容器本体
223 内蓋
223b 円錐状広がり部
225 円錐状広がり部
225a 円筒部
233b 断熱層
233f フィルムヒータ
311 円筒部 DESCRIPTION OFSYMBOLS 1 Transportation apparatus 3 Outer container 9 High specific gravity solution 11 Low specific gravity solution 13 Interface 15 Cell 17 Collection | recovery basket 19 Trap frame 21 Tube 21c Sealing part 23 Inner lid 23a Air discharge pipe 23b Conical part (conical spreading part)
23c Sealing part 31 Outer lid 33b Heat insulation part 33c Acrylic cylinder 33d Aluminum plate 33e Aluminum tape 33f Carbon 33g Film heater 33s Spring 35 Control part 35b USB
117Recovery rod 121 Tube (solution injection pipe)
121c Tube cap 123a Air vent pipe 123c Air vent cap 123d Inner cap 200 Cell storage container 201 Centrifuge tube (holding container)
213 Inner lid (container lid)
215Tissue piece 222 Container body 223 Inner lid 223b Conical spreading part 225 Conical spreading part 225a Cylindrical part 233b Heat insulation layer 233f Film heater 311 Cylindrical part
3 外側容器
9 高比重溶液
11 低比重溶液
13 界面
15 細胞
17 回収笊
19 笊枠
21 チューブ
21c 封止部
23 内蓋
23a 空気排出パイプ
23b 円錐状部(円錐状広がり部)
23c 封止部
31 外蓋
33b 断熱部
33c アクリル筒
33d アルミ板
33e アルミテープ
33f カーボン
33g フィルムヒータ
33s スプリング
35 制御部
35b USB
117 回収笊
121 チューブ(溶液注入パイプ)
121c チューブ用キャップ
123a 空気抜きパイプ
123c 空気抜き部用キャップ
123d 内蓋キャップ
200 細胞収納容器
201 遠沈管(保持容器)
213 内蓋(容器蓋)
215 組織片
222 容器本体
223 内蓋
223b 円錐状広がり部
225 円錐状広がり部
225a 円筒部
233b 断熱層
233f フィルムヒータ
311 円筒部 DESCRIPTION OF
117
213 Inner lid (container lid)
215
Claims (6)
- 細胞または組織を搬送する際に用いられる輸送装置であって、
界面が形成される高比重溶液と低比重溶液とを収容する保持容器と、
前記界面に保持される細胞または組織とは離隔して前記保持容器内に配設され、前記界面に保持された細胞または組織を回収するための回収笊と
を有することを特徴とする輸送装置。 A transport device used to transport cells or tissues,
A holding container containing a high specific gravity solution and a low specific gravity solution in which an interface is formed;
A transport apparatus, comprising: a collection basket disposed in the holding container so as to be separated from the cells or tissues held at the interface and for collecting the cells or tissues held at the interface. - 前記保持容器の上開口部を閉鎖する容器蓋と、
この容器蓋の下側に配設された空気排出孔、この空気排出孔の下端部から下方に向かって円錐状に広がり、この広がった下端の開口部が前記保持容器の内壁に近接する円錐状広がり部、およびこの円錐状広がり部の周縁部寄りに形成され、前記保持容器内に溶液を投入するための溶液注入孔を有する空気排出部と、
前記空気排出孔内に下端が挿通され、上端が上方に突出して前記容器蓋の下面に形成された円筒状中央凹部に密接に嵌合する空気排出パイプと、
上端が前記溶液注入孔を貫通して上方に突出し前記容器蓋の下面の周縁寄りに形成された円筒状周縁凹部に密接に嵌合し、下端が下方に延出し、前記回収笊を保持する溶液注入パイプと
を有することを特徴とする請求項1記載の輸送装置。 A container lid for closing the upper opening of the holding container;
An air discharge hole disposed on the lower side of the container lid, and a conical shape in which an opening at the lower end of the air expands downward in a conical shape from the lower end of the air discharge hole and is close to the inner wall of the holding container. An air discharge part formed near the peripheral part of the spreading part and the conical spreading part, and having a solution injection hole for introducing a solution into the holding container;
An air discharge pipe having a lower end inserted through the air discharge hole, and an upper end protruding upward and closely fitting into a cylindrical central recess formed on the lower surface of the container lid;
A solution that has an upper end that penetrates the solution injection hole and projects upward, closely fits into a cylindrical peripheral recess formed near the periphery of the lower surface of the container lid, and a lower end that extends downward to hold the recovery rod The transportation apparatus according to claim 1, further comprising an injection pipe. - 前記保持容器の周壁部に形成され、該保持容器内の温度を所定の温度に加温するためのヒータを有することを特徴とする請求項1または2に記載の輸送装置。 The transport apparatus according to claim 1 or 2, further comprising a heater formed on a peripheral wall portion of the holding container for heating the temperature in the holding container to a predetermined temperature.
- 前記高比重溶液は、パーフルオロカーボンまたは変性シリコーンオイルであり、前記低比重溶液は、細胞または組織の培養用の培地または細胞または組織の保存液であることを特徴とする請求項1乃至3のいずれか1項に記載の輸送装置。 4. The high specific gravity solution is perfluorocarbon or modified silicone oil, and the low specific gravity solution is a culture medium for cell or tissue culture or a cell or tissue preservation solution. The transport apparatus according to claim 1.
- 前記回収笊は、ナイロンメッシュからなる網目状部分とこの網目状部分の外周縁部を囲む笊枠で構成されることを特徴とする請求項1乃至4のいずれか1項に記載の輸送装置。 The transport device according to any one of claims 1 to 4, wherein the recovery basket includes a mesh-like portion made of a nylon mesh and a cage frame surrounding an outer peripheral edge of the mesh-like portion.
- 少なくとも前記笊枠および保持容器のいずれかの内壁にタンパク質吸着阻害剤を塗布することを特徴とする請求項5に記載の輸送装置。 6. The transport device according to claim 5, wherein a protein adsorption inhibitor is applied to at least an inner wall of either the frame or the holding container.
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