CN216337703U - Shake flask and device for continuous culture and sampling of cells - Google Patents

Abstract

The utility model discloses a shake flask and a device for continuous cell culture and sampling, wherein the shake flask for continuous cell culture and sampling comprises a flask body, wherein the flask body is provided with a bottleneck, and a first exhaust channel is formed on a first thread arranged on the bottleneck; the adapter comprises a tube body part with two through ends, the tube body part is provided with second threads matched with the first threads, the first threads and the second threads are in threaded connection to enable the adapter to be fixed on the bottle body, the outer side of the tube body part is connected with at least one interface tube, the at least one interface tube is arranged in a hollow mode and is communicated with the space in the tube body part, and third threads are arranged on the outer peripheral wall, close to the upper end opening, of the tube body part; the inner peripheral wall of the bottle cap is provided with a fourth thread matched with the third thread; and the liquid suction pipe is arranged in the bottle body, one end of the liquid suction pipe is connected with the mouth piece, and the liquid suction pipe is provided with an air vent. According to the technical scheme, the transfer and sampling of the culture liquid are performed through the adaptor, so that the risk of pollution is greatly reduced.

Description

Shake flask and device for continuous culture and sampling of cells

Technical Field

The utility model relates to the technical field of cell culture, in particular to a shake flask and a device for continuous culture and sampling of cells.

Background

The shake flask culture method is a method in which cells of a microorganism, an animal or a plant are inoculated into a shake flask containing a liquid medium, and the shake flask is placed in a shaking incubator and is continuously shaken for culture. The shake flask needs to be kept in a sealed state during the cell culture process, and is transferred to a larger-scale shake flask or reactor for amplification culture after reaching a certain cell density. In the traditional liquid transfer mode, the liquid is often directly poured into a next-level shake flask or a reactor from a small-capacity shake flask in a safety cabinet, the transfer mode has high requirements on skills of experimenters, and improper operation can cause pollution. In order to reduce the difficulty of shaking the bottle liquid transfer, some existing manufacturers adopt a mode of directly increasing an interface on a bottle cap to carry out liquid transfer, and although the liquid transfer is simple and convenient, the experience of a liquid adding user in the daily culture process is not good enough.

SUMMERY OF THE UTILITY MODEL

The utility model mainly aims to provide a shake flask for continuous cell culture and sampling, and aims to provide a convenient and flexible shake flask for cell liquid transfer in continuous cell culture.

In order to achieve the above objects, the present invention provides a shake flask for continuous culture and sampling of cells, comprising:

the exhaust device comprises a bottle body, a first exhaust channel and a second exhaust channel, wherein the bottle body is provided with a bottleneck, the upper end of the bottleneck is provided with a bottleneck, the outer peripheral wall of one end, close to the bottleneck, of the bottleneck is provided with a first thread, and the first thread is formed on the first thread;

the adapter comprises a tube body part with two through ends, a second thread matched with the first thread is arranged on the inner wall of the lower end opening of the tube body part, the first thread is in threaded connection with the second thread to enable the adapter to be fixed on the bottle body, at least one interface tube is connected to the outer side of the tube body part, the at least one interface tube is arranged in a hollow mode and communicated with the space in the tube body part, and a third thread is arranged on the outer peripheral wall, close to the upper end opening, of the tube body part;

the inner peripheral wall of the bottle cap is provided with a fourth thread matched with the third thread, and the bottle cap is fixed on the adapter through the threads; and

the pipette is arranged in the bottle body, one end of the pipette is connected with the mouth piece, and the pipette is provided with a vent hole.

Optionally, the interface tube includes a first interface section and a second interface section, the first interface section is exposed from the tube body portion, the second interface section penetrates through the tube body portion, and an outer diameter of the first interface section is larger than an outer diameter of the second interface section so that a step is formed therebetween.

Optionally, the mouthpiece is angled upwardly.

Optionally, the first thread includes a plurality of thread turns, each of the thread turns is provided with a notch, and the notches are communicated with each other to form the first exhaust passage.

Optionally, the notch is flared in a radial direction of the neck.

Optionally, the shake flask further comprises a sealing member protruding from the outer circumference of the bottle neck and disposed below the first thread.

Optionally, a second vent passage is formed on the third thread.

Optionally, a stopping portion is further disposed on an outer wall of the pipe body portion, and the stopping portion is located below the third thread.

Optionally, an opening is formed in the top end of the bottle cap, a filter membrane is arranged at the opening, and the filter membrane is used for sealing the opening.

The utility model also provides a device for continuously culturing and sampling cells, wherein the number of the mouth-piece of each of the shake flasks is two, the two mouth-pieces are oppositely arranged on the outer side of the tube body part, the capacity of the plurality of the bottle bodies is gradually increased, and one pipette is connected with one of the mouth-pieces from one of the bottle bodies; the connecting pipe comprises a liquid outlet pipe and a liquid inlet pipe, one end of the liquid outlet pipe is connected with a connector pipe connected with the liquid suction pipe from the outside, and one end of the liquid inlet pipe is connected with the other connector pipe; the other end of the liquid outlet pipe of the front-stage shake flask is fixedly communicated with the other end of the liquid inlet pipe of the rear-stage shake flask, and the liquid outlet pipe of the front-stage shake flask can be in butt joint with a power device so as to transfer liquid in the front-stage shake flask into the rear-stage shake flask.

According to the technical scheme, the bottle body is provided with a bottleneck and a bottleneck, the outer peripheral wall of one end, close to the bottleneck, of the bottleneck is provided with a first thread, the adapter comprises a pipe body part with two through ends, a second thread matched with the first thread is arranged on the inner wall of an opening at the lower end of the pipe body part, the first thread and the second thread are in threaded connection, so that the adapter is fixed on the bottle body, the pipette is arranged in the bottle body, one end of the pipette is connected with the interface tube, therefore, when the shake flask is used for cultivation, cultivation liquid in the shake flask needs to be transferred, the transfer of the cultivation liquid can be carried out through the adapter, only aseptic operation of pipeline connection is needed during liquid transfer, the risk of pollution is greatly reduced, and the overall height of the shake flask is greatly reduced because the interface tube is arranged on the side surface.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the structures shown in the drawings without creative efforts.

FIG. 1 is a schematic perspective view of a shake flask according to an embodiment of the present invention for continuous culture and sampling of cells;

FIG. 2 is a schematic perspective view of another angle of the shake flask for continuous cell culture and sampling in FIG. 1;

FIG. 3 is a schematic perspective view of the shake flask of FIG. 1 used for continuous cell culture and sampling at an angle;

FIG. 4 is a schematic exploded view of the shake flask of FIG. 3 for continuous cell culture and sampling;

FIG. 5 is a schematic perspective view of the flask of FIG. 4 for continuous cell culture and sampling;

FIG. 6 is a cross-sectional view of the bottle of FIG. 5;

FIG. 7 is a perspective view of the closure of FIG. 4;

FIG. 8 is a perspective view of the junction of FIG. 4;

fig. 9 is a perspective view of the connecting member in fig. 8.

The reference numbers illustrate:

the implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

In the present invention, unless otherwise expressly stated or limited, the terms "connected," "secured," and the like are to be construed broadly, and for example, "secured" may be a fixed connection, a removable connection, or an integral part; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.

In addition, the descriptions related to "first", "second", etc. in the present invention are only for descriptive purposes and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In addition, technical solutions between various embodiments may be combined with each other, but must be realized by a person skilled in the art, and when the technical solutions are contradictory or cannot be realized, such a combination should not be considered to exist, and is not within the protection scope of the present invention.

The flask culture method is a method in which cells of a microorganism, an animal or a plant are inoculated into a flask containing a liquid medium, and the flask is placed in a shaking incubator and cultured while shaking constantly. The shake flask needs to be kept in a sealed state during the cell culture process, and is transferred to a larger-scale shake flask or reactor for amplification culture after reaching a certain cell density. In the traditional liquid transfer mode, the liquid is often directly poured into a next-level shake flask or a reactor from a small-capacity shake flask in a safety cabinet, the transfer mode has high requirements on skills of experimenters, and improper operation can cause pollution. In order to reduce the difficulty of shaking the bottle liquid transfer, some existing manufacturers adopt a mode of directly increasing an interface on a bottle cap to carry out liquid transfer, and although the liquid transfer is simple and convenient, the experience of a liquid adding user in the daily culture process is not good enough.

To this end, the present invention provides a shake flask for continuous culture and sampling of cells.

Referring to fig. 1 to 4, in the embodiment of the present invention, the shake flask 100 for continuous cell culture and sampling includes a flask body 10, the flask body 10 has a flask neck 11, a flask mouth 12 is formed at an upper end of the flask neck 11, a first screw 111 is formed on an outer peripheral wall of one end of the flask neck 11 close to the flask mouth 12, and a first exhaust channel 112 is formed on the first screw 111; the adapter 20 comprises a tube body part 21 with two through ends, a second thread 213 matched with the first thread 111 is arranged on the inner wall of the lower end opening 41 of the tube body part 21, the first thread 111 and the second thread 213 are in threaded connection to enable the adapter 20 to be fixed on the bottle body 10, at least one interface tube 22 is connected to the outer side of the tube body part 21, the at least one interface tube 22 is arranged in a hollow mode and communicated with the space in the tube body part 21, and a third thread 211 is arranged on the outer peripheral wall, close to the upper end opening 41, of the tube body part 21; the inner peripheral wall of the bottle cap 40 is provided with a fourth thread 42 matched with the third thread 211, and the bottle cap 40 is fixed on the adapter 20 through threads; and a pipette disposed in the vial 10, one end of the pipette being connected to the mouthpiece 22, the pipette being provided with a vent hole.

In an embodiment of the present invention, the bottle body 10 may be a conical bottle. The bottle body 10 may be made of medical Polycarbonate (PC), and has the advantages of high transparency, strong impact resistance, oxidation resistance, high temperature resistance (121 ℃), and the like. A plurality of scale marks can be printed on the bottle body 10, so that an experimenter can observe the volume of the culture medium in the bottle body 10. Or PETG material is adopted. The bottle body 10 can be independently packaged in a sterile manner, so that an experimenter does not need to sterilize before using the bottle body, and the bottle body is convenient to use. The bottle body 10 may be provided with a liquid-proof portion (e.g., a protruding ring structure) at the mouth 12 thereof, so as to facilitate the pouring of the solution. The bottle neck 11 of the bottle body 10 can be set to be longer, so that the experimenter can hold the bottle neck and reduce the probability that the experimenter hands pollute the bottle neck 12.

The bottle 10 may be 125ml, 250ml, 500ml, 1500ml, 2000ml, 3000ml or 5000ml in size. The bottle body 10 with the specification of 3000ml or 5000ml has a large volume, is inconvenient to take, and a handle can be arranged on the bottle body 10 for taking conveniently. The handle may be fitted over the neck 11 of the bottle and below the bottle stopper 14. The handle may also be located on one side of the bottle 10 below the bottle stopper 14. According to different specifications, the diameter of the neck 11 can be greater than or equal to 35mm and less than or equal to 100 mm; the diameter of the bottle bottom can be greater than or equal to 65mm and less than or equal to 230 mm; the height of the bottle 10 may be greater than or equal to 110mm and less than or equal to 300 mm.

The structure of the exhaust passage has various structures, and the exhaust passage can be an exhaust notch or an exhaust opening, which will be described in detail below and will not be described again. The number of the exhaust passages may be one or more. When the bottle cap 40 of the shake flask is close to the bottle mouth 12, the bottle cap 40 gives a downward pressure to the gas near the bottle mouth 12, the gas is discharged downwards through the vent channel on the first thread 111 under the pressure and is not blocked by the first thread 111 to be gathered between two adjacent thread turns, the gas between two adjacent thread turns becomes less, so that the gas pressure inside and outside the bottle cap 40 is different, and the atmospheric pressure outside the bottle cap 40 gives a force to the bottle cap 40 close to the bottle mouth 12, so that the connection tightness of the bottle cap 40 and the bottle mouth 12 is improved.

The transition piece (see fig. 8 and 9 in combination) may be made of PP or PE semisynthetic material such as Polyetheretherketone (PEEK), Polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), or silicone elastomer, and the adaptor 20 has a cap 40 (see fig. 7) and a bottom cap 24 (see fig. 9), and the bottom cap 24 and the cap 40 may be provided with anti-slip members 50 on the outer circumferential side for convenience in closing the bottle body 10. Specifically, the anti-slip member 50 may be a non-slip strip protruding from the bottom cover 24. The quantity of antislip strip is a plurality of. The plurality of anti-slip strips may extend in the up-down direction and be arranged at intervals in the circumferential direction of the bottom cover 24. The plurality of anti-slip strips may also be arranged in a ring extending along the circumferential direction of the bottom cover 24 and spaced in the up-down direction. The anti-slip member 50 may also be a plurality of anti-slip particles protruding from the bottom cover 24, and the anti-slip particles are spaced apart from each other. The tube body part 21 of the adapter 20 is located above the bottom cover 24, the two are arranged in a communicating manner, the thickness of the bottom cover 24 is larger than that of the tube body part 21, so that the joint has better strength, the interface tube 22 is arranged at the position, close to the bottom cover 24, of the tube body part 21, one interface tube 22 can be arranged, and the interface tube 22 can be optionally assembled with an elastomer tube. Since the mouthpiece 22 penetrates the inner wall of the adaptor 20, the space inside and outside the shake flask can be communicated from the mouthpiece 22.

The inner wall of the bottom cover 24 is provided with a second thread 213 matched with the first thread 111, and the first thread 111 is provided with an exhaust passage; thus, when the bottom cap 24 is close to the bottle mouth 12, the bottom cap 24 gives a downward pressure to the gas near the bottle mouth 12, the gas is discharged downward through the vent channel on the first thread 111 under the pressure, and is not blocked by the first thread 111 and is collected between two adjacent thread turns, the gas between two adjacent thread turns becomes less, so that the gas pressure inside and outside the bottom cap 24 is different, and the atmospheric pressure outside the bottom cap 24 gives a force to the bottom cap 24 close to the bottle mouth 12, so that the connection tightness of the bottom cap 24 and the bottle mouth 12 is improved.

The pipette (not shown) can be made of medical plastic, such as medical PP, PTFE or PVC material, and the connection between the pipette and the mouthpiece 22 can be achieved by inserting, gluing or screwing, the other end is placed at or near the bottom of the bottle 10 to facilitate the liquid transfer or other operations, and the upper part of the pipette is provided with a small hole with a diameter of 0.5-2 mm.

Referring to fig. 1 to 4 and fig. 7, the shake flask further includes a bottle cap 40, the inner peripheral wall of the bottle cap 40 is provided with a fourth thread 42 matching with the third thread 211, the top end of the bottle cap 40 is provided with an opening 41, and a filter membrane is disposed at the opening 41 to close the opening 41. The number of the openings 41 may be plural, and the plural openings 41 are provided at intervals. The opening 41 may be circular, square, triangular, polygonal, kidney-shaped, or shaped. The plurality of openings 41 may be uniform in size or non-uniform in size. The filter membrane can be a PTFE filter membrane, a plurality of filter holes with the diameter of 0.2 mu m are arranged on the filter membrane, and the filter holes can prevent microorganisms from entering and exiting, so that the pollution in the culture process is avoided. Meanwhile, gas generated by metabolism of cells in the culture process can penetrate through the filter holes to exchange with the external atmosphere. The bottle cap 40 may be colored yellow, blue, red or green to allow the experimenter to distinguish the samples. The bottle cap 40 may also be a sealing cap, i.e. the bottle cap 40 is not provided with an opening 41. The bottle cap 40 may be made of a Polyethylene (PE) material, such as HDPE or LLDPE. To increase the friction, the bottle cap 40 may also be provided with anti-slip members 50 on the outer circumferential side thereof. The specific manner and structure of the anti-slip member 50 are described above and will not be described in detail herein.

The technical scheme of the utility model is that a bottle body 10, an adapter 20 and a pipette are adopted, the bottle body 10 is provided with a bottle neck 11 and a bottle mouth 12, the peripheral wall of one end of the bottle neck 11 close to the bottle mouth 12 is provided with a first thread 111, the adapter 20 comprises a tube body part 21 with two through ends, the inner wall of a lower end opening 41 of the tube body part 21 is provided with a second thread 213 matched with the first thread 111, the first thread 111 and the second thread 213 are in threaded connection to enable the adapter 20 to be fixed on the bottle body 10, the pipette is arranged in the bottle body 10, one end of the pipette is connected with a mouthpiece 22, so that under the condition that a shake flask is used for cultivation, when the culture liquid of the shake flask needs to be transferred, the transfer of the culture liquid can be carried out through the adaptor 20, only the aseptic operation of pipeline connection needs to be carried out during pipetting, the risk of pollution is greatly reduced, and the whole height of the shake flask is greatly reduced because the interface tube 22 is arranged on the side surface.

Alternatively, for the robustness of the adapter of the pipetting process, see fig. 2, 8 or 9, the interface tube 22 comprises a first interface section 221 and a second interface section 222, the first interface section 221 is exposed out of the tube body 21, the second interface section 222 penetrates through the tube body 21, and the outer diameter of the first interface section 221 is larger than that of the second interface section 222 so that a step is formed therebetween. The pipette can be inserted into the boundary between the second interface section 222 and the first interface section 221, so that the pipette is firmly connected with the second interface section 222, and liquid cannot flow out when flowing through the pipette, in addition, one end of the flexible connecting pipe can be sleeved into the interface pipe 22 and wraps the first interface section 221, and the flexible connecting pipe can be stably connected with the interface pipe 22 due to the fact that the outer diameter of the first interface section 221 is large to form a step, and pollution caused by loosening is avoided.

Further, in order to facilitate the inflow and outflow of liquid, the mouthpiece 22 is inclined upward as shown in fig. 1 to 4. The mouthpiece 22 is arranged at the mouth of the tube, such as branches of a trunk, and the inclination angle between the mouthpiece 22 and the mouth of the tube can be set at 30-75 degrees, so as to allow the cell fluid in the shake flask to flow in/out through the mouthpiece 22.

Furthermore, in order to avoid the liquid remaining in the pipeline after sampling, the upper part of the pipette is provided with a small hole of 0.5-2mm, air in the bottle can enter the pipette through the small hole, and the liquid in the pipette naturally descends under the action of gravity.

Further, referring to fig. 1 to 5, the first thread 111 includes a plurality of thread turns, each of the thread turns having a notch, and the notches are communicated with each other to form a first exhaust passage 112. In one embodiment. The shape of the notch is various, and the notch can be roughly in a semicircular shape, a triangular shape, a square shape or a trapezoidal shape. One or more notches may be provided in the same ring, without limitation.

Further, referring to fig. 1 to 5, the notch is disposed in a flaring manner in a radial direction of the bottle neck 11. With the arrangement, the gap has a larger area, and the gas is discharged from the gap at a higher speed, so that the content of residual gas between two adjacent thread turns is less, and the connection tightness between the bottom cover 24 and the bottle mouth 12 is further improved. In one embodiment, the plurality of notches are arranged in a straight line shape in the vertical direction. With the arrangement, the straight line distance between two adjacent notches is shorter, the downward gas discharge speed is higher, and the connection tightness between the bottom cover 24 and the bottle mouth 12 is further improved. Of course, the plurality of notches may be arranged in a curved shape in the up-down direction.

In another embodiment, the plurality of threaded rings of the first thread 111 may have openings formed therethrough, and the openings are communicated with each other to form an exhaust passage. The shape of the opening can be round, square, triangle, polygon, kidney or special shape. The number of the holes on the same thread turn can be one or more. The plurality of openings may be arranged in a straight line or a curved line in the up-down direction. A plurality of gaps and a plurality of holes can be formed in the plurality of threaded rings of the first thread 111, and the plurality of gaps and the plurality of holes are communicated with each other to form an exhaust channel, so that the effect of exhausting gas downwards can be achieved. It will be understood that the above arrangement may be provided in the third thread 211, that is, the outer peripheral wall of the body portion 21 near the upper end opening 41 is provided with the third thread 211, and the third thread 211 is formed with the second exhaust passage 212.

Further, referring to fig. 4 and 5, the shake flask further includes a sealing member 13, and the sealing member 13 is disposed on the periphery of the bottle neck 11 and below the first thread 111. The flask includes a sealing member 13, the sealing member 13 is protruded from the outer periphery of the neck 11 and is disposed below the first thread 111, the flask body 10 has a bottom, and the outer side surface of the sealing member 13 is inclined in a direction away from the center line of the flask body 10 in a direction in which the mouth 12 faces the bottom. In particular, the seal 13 is annular. The outward inclination of the sealing member 13 guides the movement of the cap 40 during the screwing of the cap 40 to the first thread 111, thereby preventing the bottle body 10 from falling down during the screwing. On the other hand, the cap 40 abuts the edge of the sealing member 13, enhancing the effect of the cap 40 in sealing the mouth 12.

With continued reference to fig. 4 and 5, the bottle includes a bottle stopper 14, the bottle stopper 14 is disposed protruding out of the bottle neck 11 and below the sealing member 13. The bottle stopper 14 has various structures. The bottle stopper 14 may be a stopper plate extending along the circumference of the bottle neck 11, and the bottle stopper 14 may have a circular ring shape such that the bottle stopper 14 is adapted to the shape of the bottle cap 40. The stop piece can also be a stop block which can be triangular, square or circular arc-shaped. The number of the stop blocks can be one; there may be a plurality of stoppers arranged at intervals along the circumferential direction of the bottle neck 11. The stop piece can prevent the bottle cap 40 from moving downwards continuously, and the bottle mouth 12 is prevented from being damaged by the bottle cap 40. The bottom surface of the stopper is inclined toward the center line of the bottle body 10 in the direction of the mouth 12 toward the bottom. With the arrangement, a triangular structure is formed between the top surface of the stop piece and the bottom surface of the stop piece, so that the stability is high. The stopper thus has a relatively high bearing capacity, which prevents the stopper from being broken by the pressure of the bottle cap 40. In addition, a stopper is also provided on the outer wall of the body portion 21, and the adaptor stopper 23 is located below the third thread 211. The switching element stopper is provided in the same manner as the bottle stopper 14, and will not be described here.

The present invention further provides a device for continuously culturing and sampling cells, the device for continuously culturing and sampling cells includes a connecting tube and a plurality of shake flasks 100 for continuously culturing and sampling cells, the specific structure of the shake flasks 100 for continuously culturing and sampling cells refers to the above embodiments, and the device for continuously culturing and sampling cells at least has all the advantages brought by the technical solutions of the above embodiments because all the technical solutions of the above embodiments are adopted, and details are not repeated herein.

The two mouth-pieces 22 of the shake flask are arranged oppositely outside the tube body 21, the capacities of the plurality of bottle bodies 10 are increased step by step, and a liquid suction tube is connected with one of the mouth-pieces 22 from one bottle body 10; the connecting pipe comprises a liquid outlet pipe and a liquid inlet pipe, one end of the liquid outlet pipe is connected with a connector 22 connected with a liquid suction pipe from the outside, and one end of the liquid inlet pipe is connected with the other connector 22; the other end of the liquid outlet pipe of the front-stage shake flask is fixedly communicated with the other end of the liquid inlet pipe of the rear-stage shake flask, and the liquid outlet pipe of the front-stage shake flask can be in butt joint with a power device so as to transfer liquid in the front-stage shake flask into the rear-stage shake flask. It will be appreciated that the two ports 22 are provided to accommodate the operational requirements of the transfer of the culture. In some cases, more than two mouthpiece 22 may be provided according to the use requirements, such as to facilitate measurement and the like.

Primary shake flasks refer to those selected for primary cell culture and which require transfer of cytosol from the flask.

Secondary shake flask, meaning the shake flask receiving the cell sap of the primary shake flask, and the secondary shake flask has a larger volume than the primary shake flask.

The plug represents a sealing element 13 for sealing off the connection pipe, which can be connected to the liquid inlet pipe, the liquid outlet pipe, or the mouthpiece 22.

In one case, when the cell concentration in the cell culture fluid in the primary flask is increased to a certain degree, a secondary flask may be selected for pipetting, specifically, the plugs at both ends of the adaptor 20 of the primary flask are removed, the liquid inlet pipe and the liquid outlet pipe are connected to the liquid inlet of the primary flask, the liquid inlet pipe is connected to the liquid outlet pipe of the secondary flask, the liquid outlet pipe of the primary flask and the liquid inlet pipe of the secondary flask are connected, and the liquid in the primary flask is transferred to the secondary flask by a peristaltic pump or a compressed air power device. So, only need carry out the aseptic technique of tube coupling when shaking the bottle to elementary and move the liquid, the risk of greatly reduced pollution to need not to take out the pipette wholly when shaking the bottle and uncapping in the culture, only need with bottle lid 40 open can, avoid the pipette to contact the mesa and cause the pollution, user daily operation is not influenced.

When the shake flask needs to be sampled, the shake flask needing to be sampled is selected, a clean disposable injection molding device is prepared, the plug on the liquid outlet is taken down, the injector is connected to the joint of the liquid outlet, after appropriate liquid is extracted, the injector is taken down, and the plug is connected back to the liquid outlet. In order to avoid the liquid remaining in the pipeline after sampling, a pipette is placed in the bottle body, one end of the pipette is connected with the mouth piece 22, and the pipette is provided with a vent hole (not marked in the figure). The vent hole is a small hole with the diameter of 0.5-2mm arranged at the upper part of the pipette, air in the bottle can enter the pipette through the small hole, and liquid in the pipette naturally descends under the action of gravity.

In another case, when high-density cell culture is required, liquid supplement and aeration can be performed through a liquid outlet, so that continuous long-time culture is realized. Specifically, the plug of the liquid outlet is taken down and connected with a three-way joint, one end of the three-way joint is connected with an air inlet filter, and the other end of the three-way joint is connected with a liquid inlet bottle. Adjusting the air inlet pressure and flow of the device to make the filtered air flow out from the bottom of the shake flask, clamping the hose into a peristaltic pump, setting a proper rotating speed, and starting material supplement. Furthermore, parallel culture with the shake flasks described herein mounted on a shaking platform in one incubator significantly reduces the culture cost and complexity, thus still allowing faster, more efficient and cost-reduced process development under reproducible and comparable conditions.

The above description is only an alternative embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications and equivalents of the present invention, which are made by the contents of the present specification and the accompanying drawings, or directly/indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims (10)

1. A shake flask for continuous culture and sampling of cells, comprising:

the exhaust device comprises a bottle body, a first exhaust channel and a second exhaust channel, wherein the bottle body is provided with a bottleneck, the upper end of the bottleneck is provided with a bottleneck, the outer peripheral wall of one end, close to the bottleneck, of the bottleneck is provided with a first thread, and the first thread is formed on the first thread;

the adapter comprises a tube body part with two through ends, a second thread matched with the first thread is arranged on the inner wall of the lower end opening of the tube body part, the first thread is in threaded connection with the second thread to enable the adapter to be fixed on the bottle body, at least one interface tube is connected to the outer side of the tube body part, the at least one interface tube is arranged in a hollow mode and communicated with the space in the tube body part, and a third thread is arranged on the outer peripheral wall, close to the upper end opening, of the tube body part;

the inner peripheral wall of the bottle cap is provided with a fourth thread matched with the third thread, and the bottle cap is fixed on the adapter through the threads; and

the pipette is arranged in the bottle body, one end of the pipette is connected with the mouth piece, and the pipette is provided with a vent hole.

2. A shake flask for continuous culture and sampling of cells according to claim 1, wherein the mouthpiece comprises a first interface section and a second interface section, the first interface section being exposed to the body portion, the second interface section penetrating the body portion, the first interface section having an outer diameter larger than the second interface section such that a step is formed therebetween.

3. A shake flask for continuous culture and sampling of cells according to claim 2, wherein the mouthpiece is tilted upwards.

4. A shake flask for continuous culture and sampling of cells according to claim 1, wherein the first screw thread comprises a plurality of turns, each turn having a notch, the notches communicating with each other to form the first vent channel.

5. A shake flask for continuous culture and sampling of cells according to claim 4, wherein the notch is flared in the radial direction of the neck.

6. A shake flask for continuous culture and sampling of cells according to claim 1, further comprising a sealing member protruding from the outer circumference of the neck and arranged below the first thread.

7. A shake flask for continuous culture and sampling of cells according to claim 1, wherein the third screw thread is formed with a second degassing channel.

8. A roller bottle according to claim 7, wherein the outer wall of the body is further provided with a stop, and the stop is located below the third thread.

9. A shake flask for continuous cell culture and sampling according to claim 7, wherein the top of the bottle cap is provided with an opening, and the opening is provided with a filter membrane for closing the opening.

10. An apparatus for continuous cell culture and sampling, comprising:

a plurality of shake flasks as defined in any one of claims 1 to 9, wherein the number of the mouth-piece tubes is two, two of the mouth-piece tubes are oppositely arranged outside the tube body part, the capacity of the plurality of the flask bodies is gradually increased, and a pipette is connected with one of the mouth-piece tubes from inside one of the flask bodies; and

the connecting pipe comprises a liquid outlet pipe and a liquid inlet pipe, one end of the liquid outlet pipe is connected with a connector pipe connected with the liquid suction pipe from the outside, and one end of the liquid inlet pipe is connected with the other connector pipe;

the other end of the liquid outlet pipe of the front-stage shake flask is fixedly communicated with the other end of the liquid inlet pipe of the rear-stage shake flask, and the liquid outlet pipe of the front-stage shake flask can be in butt joint with a power device so as to transfer liquid in the front-stage shake flask into the rear-stage shake flask.

Images