JPWO2017163326A1 - Cultivation container and automatic culture apparatus using the same - Google Patents

Abstract

Provided are a culture vessel having a simple and small flow path configuration connected to the vessel, and an automatic culture apparatus using the same. The culture container 101 includes a container 1 having a culture space inside, a container lid member 9 that seals the container, and a first container provided on the container lid member that enables fluid movement from the outside of the container to the container. It has the flow path 4 and the 2nd flow path 6 branched from the 1st flow path. The first channel can be connected to a channel outside the culture vessel, and a check valve 5 is provided between the branch position 4B and the end 4A on the vessel side to allow fluid from inside the vessel to flow. The second flow path that branches from the first flow path is provided with a check valve 7 and can flow into the container. Can be allowed to pass through, and the flow of fluid from within the container can be stopped.

Description

  The present invention relates to a culture container and an automatic culture apparatus using the same.

  Conventionally, in culture of various cells (floating cells, adherent cells), a medium culture solution containing nutrients is regularly exchanged. In particular, for culturing rapidly metabolized cells, it is necessary to exchange the culture medium at a high frequency. Conventionally, cell culture operations including such medium exchange have been performed by a skilled operator using a pipette. However, there has been a problem that cell culture in a large number of culture containers requires much labor. Recently, the development of an automated system for culture medium replacement work and culture work has been actively conducted. Among them, in particular, an automatic culture configuration using a closed culture vessel and a closed flow channel capable of maintaining sterility is attracting attention (Patent Document 1).

WO2014 / 049701 Publication

  In the configuration of Patent Document 1, when culturing with an automatic culture apparatus, it is necessary to supply and discharge a culture solution, mixed gas, and the like through channels connected to the culture vessel from the outside of the culture vessel via connection portions. Therefore, when a plurality of culture containers are used for cell culture at the same time, the configuration of the flow path connected to the culture containers is complicated, and the work of connecting the flow paths to the culture container becomes complicated, which may increase the risk of work errors. is there.

  An object of the present invention is to provide a culture vessel having a simple and small channel configuration connected to the culture vessel and an automatic culture apparatus using the same.

  In order to achieve the above object, in the present invention, a culture vessel that has a culture space inside, a vessel lid member that seals the vessel, and fluid movement from the outside to the vessel is enabled. A first flow path provided in the container lid member and a second flow path branched from the branch position of the first flow path, the first flow path being connectable to an external flow path And the first valve is provided between the branch position and the end of the first flow path on the container side to allow fluid from inside the container to pass therethrough. The second flow path has a second valve, and the second valve can pass fluid into the container, and the container can stop the flow into the container. A culture vessel that can stop the flow from inside is provided.

  In order to achieve the above object, according to the present invention, there is provided an automatic culture apparatus, which is at least one culture container, a first storage part for storing a fluid discharged from the culture container, A second container that contains the fluid to be supplied to, a flow path outside the culture container that is branched from the culture container and connected to the first container and the second container, and outside the culture container An open / close valve provided in the flow path for controlling the flow of the fluid, a pump for supplying and discharging the fluid to and from the culture vessel, and an open / close valve and a controller for controlling the operation of the pump, The culture container includes a container having a culture space therein, a container lid member for sealing the container, and a first flow path provided in the container lid member that enables fluid movement from the outside of the culture container to the container. And a second flow path that branches from the branch position of the first flow path, The first flow path has a connection portion and a first valve connectable to a flow path outside the culture vessel, and the first valve has a branch position and an end on the container side of the first flow path. The fluid from the inside of the container can be passed, the flow of the fluid into the container can be stopped, the second flow path has a second valve, The second valve provides an automatic culture device that can pass fluid into the container and stop the flow of fluid from the container.

  According to the present invention, the flow path configuration connected to the culture vessel is simple and small.

FIG. 3 is a diagram illustrating a configuration example of a culture vessel according to Example 1. 1 is a diagram illustrating a schematic configuration example of an automatic culture apparatus according to Example 1. FIG. 1 is a diagram illustrating a configuration example of a check valve according to Embodiment 1. FIG. It is a figure which shows the flow at the time of the culture solution supply of the automatic culture apparatus which concerns on Example 1. FIG. It is a figure which shows the flow at the time of the culture solution discharge | emission of the automatic culture apparatus which concerns on Example 1. FIG. FIG. 3 is a diagram showing a flow of mixed gas supply and exhaust of the automatic culture apparatus according to Example 1. FIG. 3 is a diagram showing a configuration example of simultaneous culture of a plurality of culture vessels according to Example 1. FIG. 4 is a diagram illustrating a configuration example of a culture vessel according to Example 2. It is a figure which shows the flow at the time of the culture solution supply of the automatic culture apparatus using the culture container which concerns on Example 3. FIG.

  Hereinafter, various embodiments of the present invention will be described with reference to the drawings. In the present specification, gas, liquid, gas and liquid flowing through the flow path of the automatic culture apparatus may be collectively referred to as fluid. In this specification, a valve is a general term for devices having a mechanism capable of opening and closing a fluid passage, and the term “valve” may be used, and includes, for example, an on-off valve and a check valve. Furthermore, in this specification, the container means a container itself having a cell culture space inside, and the culture container includes the container, a container lid member, a flow path that enables fluid movement from the outside of the container, and the like. It shall mean a structure. Concerning the numbers of the components on the drawings, the numbers already described in other drawings may be omitted in other drawings. Moreover, about the one part figure which shows an automatic culture apparatus, the figure of the state which abbreviate | omitted the temperature control unit etc. which control the temperature of an incubator, the control terminal which comprises a control part, an incubator, etc. shall be shown.

  Example 1 includes a container having a culture space inside, a container lid member for sealing the container, and a first flow path provided in the container lid member that enables fluid movement from the outside with respect to the container. A second flow path that branches from the branch position of the first flow path, the first flow path having a connection portion that can be connected to an external flow path and a first valve, The valve is provided between the branch position and the container-side end of the first flow path, can pass the fluid from inside the container, and can stop the flow into the container. The second flow path has a second valve, and the second valve allows the fluid into the container to pass therethrough and stops the flow from within the container; and It is an Example of the automatic culture apparatus using it.

  FIG. 1A is a diagram showing a configuration of a culture vessel according to Example 1, and FIG. 1B is a diagram showing a schematic configuration of an automatic culture apparatus using the culture vessel according to Example 1.

  First, an example of the structure of the culture vessel according to Example 1 will be described with reference to FIG. 1A. The culture container 101 includes a container 1 as a first container having a culture space for cells 3 therein, a container lid member 9 that seals the container 1, and fluid movement of a culture solution or a mixed gas from the outside of the container 1 The first flow path 4 provided in the container lid member 9 and the second flow path 6 branched from the first flow path 4 at the branch position 4B are provided. As shown in the figure, the second channel 6 is shorter than the first channel 4, and the second channel 6 has a length that does not reach the culture solution 2 held in the container 1. The channel 4 has a length that reaches the culture solution 2 in the container 1 when the culture container 101 is held at a predetermined angle.

  The first flow path 4 has a check valve 5 that is a first valve in the flow path, and the second flow path 6 is a check that is a second valve in the flow path. It has a valve 7. The check valve 5 of the first channel 4 is provided between the branch position 4B and the end 4A of the first channel 4 on the container 1 side. The check valve 7 of the second flow path 6 is provided between the branch position 4 </ b> B from the first flow path 4 and the container-side end 6 </ b> A of the second flow path 6.

  Here, the direction in which the fluid can pass through the check valves 5 and 7 shown in the figure follows the direction of the arrow shown in FIG. That is, the check valve 5 of the first flow path 4 is installed so that the fluid from the inside of the container 1 can pass and the flow of the fluid into the container 1 can be stopped. The check valve 7 of the path 6 is installed so that the fluid into the container 1 can pass and the flow of the fluid from the container 1 can be stopped. In the check valves other than the check valves 5 and 7 shown in the other drawings, the direction in which the fluid can pass is the direction shown in FIG.

  The position of the first flow path 4 in the container 1 is such that the end 4A on the container 1 side of the first flow path 4 is at the lowest point of the container 1 when the culture solution 2 accommodated in the container 1 is discharged. It is desirable to install so that it is located in. By doing so, when the culture vessel 101 is held at an arbitrary angle, the culture solution 2 gathers at the lowest point of the vessel 1, so that the discharge efficiency of the culture solution 2 can be improved.

  The position of the second flow path 6 in the container 1 should be changed depending on the amount of the culture medium 2 accommodated in the container 1, but the length of the flow path that does not reach the culture liquid 2 held in the container 1. By doing so, it is only necessary to be above the liquid level of the contained culture solution 2. Thus, the first channel 4 and the second channel 6 can be arranged at different positions in the container 1 by the configuration in which the second channel 6 is branched from the first channel 4. is there.

  The first flow path 4 has a sterile connection portion 8 that can be aseptically connected to a flow path outside the culture vessel 101. The container 1 is provided with an exhaust port 10 for discharging the gas in the container 1 to the outside of the culture container 101. The exhaust port 10 is provided with an air filter 11 so that microorganisms and the like do not enter from the outside of the culture vessel 101 when the gas passes.

  Next, a configuration example of an automatic culture apparatus using the culture container according to Example 1 will be described with reference to FIG. 1B. The automatic culture apparatus 102 using the culture container 101 is supplied into the culture container 101 and the drainage bag 15, which is a first storage unit that stores the culture liquid 16 that is a liquid discharged from the culture container 101. A liquid supply bag 18 that is a second storage unit that stores a culture solution 19 that is a liquid to be supplied, and an air supply device 24 for supplying a mixed gas that is a gas necessary for culture into the culture vessel.

  The flow path 13 outside the culture container has a sterile connection part 12 and is connected to the culture container 101 in combination with the sterile connection part 8 provided in the first flow path 4. The channel 13 outside the culture vessel is branched and connected from the culture vessel 101 to the drainage bag 15, the liquid supply bag 18, and the air supply device 24. Solenoid valves 17, 20, and 26 are provided as opening and closing valves that can be opened and closed.

  An air filter 25 is provided in the flow path between the air supply device 24 and the electromagnetic valve 26. The flow path 13 outside the culture vessel is further provided with a pump 14, an air supply port 23 having an air filter 22, and an electromagnetic valve 21 as an open / close valve that allows the air supply port 23 to be opened and closed. The pump 14 installed in the flow path 13 outside the culture container enables supply and discharge of fluid and gas to and from the container 1 through the flow path 13 outside the culture container.

  Since the culture vessel 101 needs to be maintained at an optimal culture temperature for cell culture, the temperature can be adjusted by the temperature control unit 28 installed in the incubator 27 and installed outside the incubator 27.

  The control terminal 30 constituting the control unit is a personal computer (PC) having a computer configuration including a normal central processing unit (CPU), a storage unit, an input / output interface unit, and the like. It is possible to control the pump 14, the solenoid valves 17, 20, 21, and 26, the temperature control unit 28, and the like via 29.

  Hereinafter, the procedure for exchanging the culture solution in the culture vessel 101 of the automatic culture apparatus 102 of this embodiment will be described with reference to FIGS. 3A, 3B, and 3C. First, as shown in FIG. 3A, when the culture solution 2 is discharged from the container 1, only the electromagnetic valve 17 is opened. When the pump 14 is operated, the culture solution 2 is discharged from the container 1 through the check valve 5 in the first flow path 4, the aseptic connection parts 8 and 12, the flow path 13 outside the culture container, and the open electromagnetic valve 17. Drain into the bag 15.

  Here, since the check valve 7 of the second flow path 6 is configured to stop the flow of fluid from the inside of the container 1, the second flow path 6 is closed by the check valve 7. Therefore, it is possible to prevent the fluid from flowing backward from the second channel 6 and to efficiently recover the culture solution 2 without reducing the pressure in the channel.

  Next, as shown in FIG. 3B, when supplying the culture solution 19, the electromagnetic valve 17 is closed and the electromagnetic valve 20 is opened. The container is opened through the solenoid valve 20 in which the culture solution 19 is opened from the supply bag 18 by the operation of the pump 14, the flow path 13 outside the culture container, the aseptic connection parts 12 and 8, and the check valve 7 of the second flow path 6. 1 is supplied.

  Here, by opening the electromagnetic valve 21 while the pump 14 is operating, the gas is introduced into the flow path 13 outside the culture vessel through the air filter 22 into the flow path 13 outside the culture vessel, and inside the flow path 13 outside the culture vessel. It is also possible to sufficiently supply the culture solution remaining in the container 1 to the container 1.

  Finally, as shown in FIG. 3C, the mixed gas necessary for cell culture is such that only the electromagnetic valve 26, which is an open / close valve, is opened, and the mixed gas is supplied from the air supply device 24 to the air filter 25, and the opened electromagnetic valve 26. Then, the liquid is supplied to the container 1 through the flow path 13 outside the culture container, the aseptic connections 12 and 8, and the check valve 7 of the medium supply flow path 6, and is discharged from the container 1 through the exhaust port 10 and the air filter 11 of the container 1. .

  In the configuration of the present embodiment, the first flow path 4 and the check valve 7 provided in the first flow path 4 and the second flow path 6 provided in the culture vessel 101 are used as the first flow path. 4, the culture solution 2 can be discharged from the container 1, and the culture solution 19 and the mixed gas can be supplied to the container 1 from the second channel 6 branched from the first channel 4. And That is, the check valve 5 that is the first valve can pass the liquid discharged from the container 1 to the drainage bag 15 that is the first container, and the liquid supply that is the second container. The check valve 7, which is a second valve, can stop the flow of the liquid supplied from the bag 18 to the container 1 and the gas supplied from the air supply device 24. The liquid supplied from the container 1 to the container 1 and the gas supplied from the air supply device 24 can pass therethrough, and the flow of the liquid discharged from the container 1 to the drainage bag 15 can be stopped.

  Thus, according to the present embodiment, the number of connection portions provided in the culture vessel can be reduced by sharing the flow path connected to the culture vessel for supplying and discharging the culture solution and supplying the mixed gas. The flow path configuration connected to the culture vessel is simple and small.

  The automatic culture apparatus of the present embodiment has mechanisms for supplying and discharging the culture solution, supplying and exhausting the mixed gas, but if a similar channel is added to the channel outside the culture vessel, the other For example, cell seeding, channel washing, introduction of a cell detachment enzyme, collection of a cell suspension, and the like can be performed.

  In the above description, the configuration of the automatic culture apparatus equipped with one culture vessel 101 is shown and described. However, a plurality of on-off valves connected to the connection portions of the plurality of culture vessels 101 are provided outside the culture vessel. It can also be set as the structure of the automatic culture apparatus which has a branch part connected to this flow path. That is, as in another configuration example shown in FIG. 4, a plurality of culture vessels 101 are mounted, and the respective culture vessels 101 are connected via the aseptic connection portions 8 and 12, the electromagnetic valve 32 that is an on-off valve, and the branch portion 31. Is connected to the flow path 13 outside the culture vessel, and the control terminal 30 sequentially controls the flow of the fluid by executing switching control of the electromagnetic valves 17, 20, 21, 26, 32 and the pump 14 as opening / closing valves. Simultaneously, the supply and discharge of the culture solution to the plurality of culture vessels 101, the supply and exhaust of the mixed gas can be performed.

  For the culture container of this embodiment, it is desirable to use a material that can withstand high humidity and sterilization, such as resin. Thereby, it can be adapted for sterilization such as gamma rays, and can be applied to physics and chemistry use and regenerative medicine use.

  1 to 4 of the present embodiment, the flow path 4 has been described in the vertical direction, but the present invention can also be applied to a horizontal direction or an oblique direction flow path. The position of the second flow path 6 in the container 1 should be changed according to the amount of the culture solution 2 accommodated in the container 1. For example, a position sensor that measures the position of the culture solution surface is not shown. Is provided in the automatic culture apparatus 102, and the supply amount of the culture solution 19 is adjusted by the control terminal 30 based on the position information of the position sensor, so that the position of the second flow path 6 is located above the culture solution 2. It is also possible to do.

  Although not shown, a cell image captured by a microscope is input to the control terminal 30 or transmitted from the control terminal 30 to the data server via a network, and the cell image is processed by various image processing software. Can also be implemented.

  The culture vessel according to Example 1 and the automatic culture apparatus using the same can be applied to two-layer culture by changing a part of the configuration. As Example 2, an example of a culture container having a two-layer structure applied to two-layer culture and an automatic culture apparatus will be described. That is, in addition to the configuration of Example 1, Example 2 is an upper layer container that is accommodated and held in a container and has a culture space inside, and a container lid member that enables fluid movement from the outside with respect to the upper layer container. A third flow path provided, and a fourth flow path branched from the branch position of the third flow path, and the third flow path includes a connection portion connectable to an external flow path. The third valve is provided between the branch position of the third flow path and the end on the upper container side of the third flow path, and is provided from the upper container. The fluid can pass therethrough and the flow of the fluid into the upper layer container can be stopped. The fourth flow path has a fourth valve, and the fourth valve is connected to the upper layer container. A culture vessel capable of allowing fluid to pass therethrough and stopping the flow of fluid from within the upper layer vessel, and using the same It is an example of a dynamic culture device.

  ES (Embryonic Stem) cells, iPS (Induced Pluripotent Stem) cells, and cell types such as skin epithelial cells, corneal epithelial cells, and oral mucosal epithelial cells, which are attracting attention in regenerative medicine, play a role in providing nutrients during culture When co-culturing feeder cells, it is desirable to culture the feeder cells and cells used for treatment in a separated state, and it is desirable to perform two-layer culture in a two-layer culture vessel.

   FIG. 5 is a view showing a culture container having a two-layer structure according to Example 2. FIG. In the figure, a two-layer culture vessel 103 includes an upper layer vessel 33, a feeder cell 34, a third channel 35, and a third channel 35 as a second vessel having a culture space for cells 3 therein. A check valve 36, which is a third valve, a fourth flow path 37, a check valve 38 which is a fourth valve of the fourth flow path 37, and a sterile connection 39 are provided. Since other parts have the same configuration as that of the first embodiment, description thereof is omitted here.

  In the culture container 103 having a two-layer structure, the container 1 having a culture space for the cells 34 therein accommodates and holds an upper layer container 33 having a culture space inside. In order to enable fluid movement of the culture solution or the like from the outside of the container 1 with respect to the upper layer container 33, a third flow path 35 and a fourth flow path branched from the third flow path are provided in the container lid member 9. 37 is provided.

  The third flow path 35 has a check valve 36 in the flow path, and the fourth flow path 37 has a check valve 38 in the flow path. As with the check valve 5 of the first flow path 4, the check valve 36 of the third flow path 35 includes the branch position of the third flow path 35 and the end on the upper container side of the third flow path 35. It is provided between the part 35A.

  As described with reference to FIG. 2, the check valve 36 of the third flow path 35 can pass the fluid from the upper layer container 33 and can stop the flow of the fluid into the upper layer container 33. The check valve 38 of the fourth flow path 37 is installed so that the fluid can be passed into the upper layer container 33 and the flow of the fluid from the upper layer container 33 can be stopped.

  The position of the third flow path 35 in the upper layer container 33 is such that when the culture solution 2 accommodated in the upper layer container 33 is discharged, the end 35A on the upper layer container side of the third flow path 35 is located on the upper layer container 33. It is desirable to install it at the lowest point. By doing so, when the culture vessel 103 having a two-layer structure is held at an arbitrary angle, the culture solution 2 collects at the lowest point of the upper layer vessel 33, so that the discharge efficiency of the culture solution 2 can be improved.

  The position of the fourth flow path 37 in the upper layer container 33 should be changed depending on the amount of the culture solution 2 accommodated in the upper layer container 33, but if it is above the liquid level of the accommodated culture solution 2 Good. The third flow path 35 has a sterile connection 39 that can be aseptically connected to a flow path outside the culture vessel.

  The two-layer culture container 103 of this embodiment is adapted to the automatic culture apparatus described in the first embodiment, and is performed in the upper container 33 for culturing the cells 3 in the first embodiment. And by exchanging the culture medium 2 in the same manner as in Example 1 in each of the container 1 and the upper layer container 33, two-layer co-culture using feeder cells becomes possible.

  Note that the two-layered culture container 103 of the present embodiment is not limited to performing two-layer co-culture using feeder cells, and different cell tumors are cultured in the container 1 and the upper container 33, respectively. It is also possible to culture the same cell tumor.

  The configuration of this example also has the same effect as that of the single-layer culture of Example 1 above. That is, the number of connection portions provided in the two-layer structure culture vessel can be reduced, and the flow path configuration connected to the culture vessel can be simplified and reduced in size.

  When bubbles are generated in the culture solution when feeding the culture solution into the culture vessel, the impact on the cells due to the impact of the bursting of the bubbles and the protein structure in the culture solution change due to the bubbles being hydrophobic. This may have an effect on the culture solution. Therefore, a device for reducing the generation of bubbles may be required.

  Example 3 is an example of a culture vessel capable of reducing the generation of bubbles in a culture solution and an automatic culture apparatus using the same in addition to the configuration of Example 1. That is, in addition to the configuration of Example 1, a culture vessel in which the container-side end of the second flow channel branched from the first flow channel is provided close to the inner wall of the container, and automatic culture using the same It is the Example of an apparatus.

  In Example 1, as shown in FIG. 1, the second flow path 6 for supplying the culture solution 19 to the container 1 has a shorter shape than the first flow path 4 and is above the liquid level of the culture liquid 2. By installing, it is possible to suppress the generation of bubbles in the culture solution when supplying the culture solution or mixed gas. Example 3 further provides a culture vessel configuration suitable for reducing the generation of bubbles during the supply of the culture solution.

  As shown in FIG. 6, as a further contrivance of the shape of the second flow path, the second flow path 6 is branched from the branch position 4B of the first flow path 4 toward the inner wall of the container 1 and then discharged. In order to make the fluid to flow along the inner wall of the container 1, the container-side end 6 </ b> A of the second flow path 6 is installed close to the inner wall of the container 1. “Installing close to the inner wall” means that the container-side end 6A of the second flow path 6 is set close to a distance along which the fluid flowing out from the second flow path 6 runs along the inner wall of the container. Means.

  With the configuration of the present embodiment, it is possible to reduce the generation of bubbles in the culture solution 2 due to the momentum of the flow when the culture solution 19 is supplied. By adopting the configuration of Example 3, as in Example 1, the configuration of the flow path connected to the culture vessel becomes simple and small, and furthermore, the influence of bubbles on the cells and the culture solution when supplying the culture solution is reduced. Can do. The configuration in which the container-side end 6A of the second flow path described in the present embodiment is close to the inner wall of the container 1 is the same as the inner wall of the upper-layer container 33 and the upper-layer container side of the fourth flow path in the second embodiment. The present invention can also be applied to the positional relationship with the end portion 37A.

  According to the present embodiment, it is possible to provide a culture vessel capable of reducing the generation of bubbles in the culture solution and reducing the influence on the cells and the culture solution, and an automatic cell apparatus using the same.

  Although various embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments unless the characteristics of the present invention are impaired, and can be considered within the scope of the technical idea of the present invention. Various other modifications are included. For example, in each of the above-described embodiments, the check valve is exemplified and described as the first and second valves. However, in place of the check valve, fluid from inside the container can be allowed to pass through the check valve. Any other structure can be used as long as the flow of fluid into the container can be stopped, or the fluid can be passed into the container and can be stopped. Can be used. The same applies to the third and fourth valves.

  Further, the above-described embodiments have been described in detail for better understanding of the present invention, and are not necessarily limited to those having all the configurations described above. Further, a part of the configuration of one embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of one embodiment. Further, it is possible to add, delete, and replace other configurations for a part of the configuration of each embodiment.

  Further, the above-described configuration, function, control unit, and the like have been described as an example of creating a CPU program that realizes part or all of them. Needless to say, it may be realized by hardware. That is, all or a part of the functions of the control unit may be realized by an integrated circuit such as an application specific integrated circuit (ASIC) or a field programmable gate array (FPGA) instead of the program.

DESCRIPTION OF SYMBOLS 1 Container 2, 16, 19 Culture solution 3 Cell 4 1st flow path 4A End part 4B of the 1st flow path on the container side Position 5, 7, 36, 38 of 1st flow path branch Check valve 6 Second flow path 6A Container side ends 8, 12, 39 of the second flow path 9 Aseptic connection 9 Container lid member 10 Exhaust port 11, 22, 25 Air filter 13 Flow path 14 outside culture vessel Pump 15 Exhaust Liquid bag 17, 20, 21, 26, 32 Solenoid valve 18 Liquid supply bag 23 Air supply port 24 Air supply device 27 Incubator 28 Temperature control unit 29 Electric signal line 30 Control terminal 31 Branching section 33 Upper layer container 34 Feeder cell 35 Third flow path 35A Third flow path upper-layer container end 37 Fourth flow path 37A Fourth flow path upper-layer container end 101 Culture container 102 Automatic culture apparatus 103 Two-layer culture container

Claims (15)

A culture vessel,
A container having a culture space inside;
A container lid member for sealing the container;
A first flow path provided in the container lid member that enables fluid movement from the outside with respect to the container;
A second flow path that branches off from the branch position of the first flow path,
The first flow path has a connection portion and a first valve connectable to an external flow path, and the first valve is located on the container side of the branch position and the first flow path. Provided between the end portion, the fluid from the inside of the container can pass therethrough, the flow of fluid into the container can be stopped,
The second flow path has a second valve, and the second valve can pass the fluid into the container and can stop the flow of fluid from the container. Is,
A culture vessel characterized by that. The culture container according to claim 1,
The first valve and the second valve are check valves, respectively.
A culture vessel characterized by that. The culture container according to claim 1,
The connection portion can aseptically connect the external flow path to the container.
A culture vessel characterized by that. The culture container according to claim 1,
The container has an exhaust port;
A culture vessel characterized by that. The culture container according to claim 1,
The container-side end of the second flow path branched from the first flow path is provided close to the inner wall of the container.
A culture vessel characterized by that. The culture container according to claim 1,
An upper layer container housed and held in the container and having a culture space inside;
A third flow path provided in the container lid member that enables fluid movement from the outside with respect to the upper layer container;
A fourth flow path branched from the branch position of the third flow path,
The third flow path has a connection portion connectable to the external flow path and a third valve,
The third valve is provided between a branch position of the third flow path and an end portion of the third flow path on the upper container side, and passes the fluid from the upper layer container. The flow of fluid into the upper layer container can be stopped,
The fourth flow path has a fourth valve, and the fourth valve can pass the fluid into the upper layer container and can stop the flow of fluid from the upper layer container. Is possible,
A culture vessel characterized by that. The culture container according to claim 6,
The third valve and the fourth valve are check valves.
A culture vessel characterized by that. The culture container according to claim 6,
The connection portion of the third flow path can aseptically connect the external flow path to the upper layer container.
A culture vessel characterized by that. The culture container according to claim 6,
An end portion on the upper layer container side of the fourth flow channel branched from the third flow channel is provided close to the inner wall of the upper layer container,
A culture vessel characterized by that. An automatic culture device,
At least one culture vessel;
A first accommodating portion for accommodating a fluid discharged from the culture vessel;
A second accommodating portion for accommodating a fluid to be supplied into the culture vessel;
From the culture container, a flow path outside the culture container that is branched and connected to the first storage part and the second storage part,
An open / close valve for controlling the flow of fluid, provided in a flow path outside the culture vessel;
A pump for supplying and discharging fluid to and from the culture vessel;
A control unit for controlling the operation of the on-off valve and the pump,
The culture vessel is
A container having a culture space inside;
A container lid member for sealing the container;
Enabling fluid movement from the outside of the culture vessel with respect to the vessel, a first flow path provided in the vessel lid member;
A second flow path that branches off from the branch position of the first flow path,
The first flow path has a connection portion and a first valve connectable to a flow path outside the culture vessel, and the first valve is connected to the branch position and the first flow path. Provided between the container and the end on the container side, the fluid from inside the container can pass therethrough, and the flow of fluid into the container can be stopped,
The second flow path has a second valve, and the second valve can pass the fluid into the container and can stop the flow of fluid from the container. Is,
An automatic culture apparatus characterized by that. The automatic culture apparatus according to claim 10, wherein
An air supply device for supplying air to the culture vessel;
The flow path outside the culture vessel is branched and connected to the air supply device,
The container includes an exhaust port;
An automatic culture apparatus characterized by that. The automatic culture apparatus according to claim 10, wherein
The first valve can pass the liquid discharged from the container to the first storage part, and can stop the flow of liquid supplied from the second storage part to the container. And
The second valve can pass the liquid supplied from the second container to the container, and can stop the flow of liquid discharged from the container to the first container. Is,
An automatic culture apparatus characterized by that. The automatic culture apparatus according to claim 10, wherein
The first valve can pass the liquid discharged from the container to the first container, and is supplied from the second container to the container and supplied from the air supply device. Can stop the flow of gas
The second valve can pass the liquid supplied from the second container to the container and the gas supplied from the air supply device, and is discharged from the container to the first container. That can stop the flow of liquid
An automatic culture apparatus characterized by that. The automatic culture apparatus according to claim 10, wherein
An incubator accommodated in the culture vessel, and a temperature control unit for controlling the temperature in the incubator,
An automatic culture apparatus characterized by that. The automatic culture apparatus according to claim 10, wherein
A plurality of on-off valves connected to each of the connection portions of the plurality of culture vessels, further comprising a branch portion for connecting to a flow path outside the culture vessel;
An automatic culture apparatus characterized by that.

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