Culture systems and cell culture methods

Abstract

To provide a culture system and a cell culture method capable of culturing cells in parallel.SOLUTION: A culture system comprising a cell container 10 configured to contain cells, a plurality of culture devices 20, and a distribution channel 30 configured to distribute the cells from the cell container 10 to the plurality of culture devices 20, where: each of the plurality of culture devices 20 comprises a plurality of culture tanks 21, a supply channel 22 configured to selectively supply fluid from the distribution channel 30 to the plurality of culture tanks 21, and a connecting channel 23 that connects the plurality of culture tanks 21; in each of the plurality of culture devices 20 the plurality of culture tanks 21 are connected in series via the connecting channel 23; and in each of the plurality of culture devices 20, the supply channel 22 selectively supplies fluid to the plurality of culture tanks 21 via the connecting channel 23 so that the cells sequentially move through the plurality of culture tanks 21.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a culture system and a method for culturing cells.

Background Art

[0002] Systems for automatically culturing cells have been proposed (see, for example, Patent Documents 1 and 2).

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0004] One object of the present invention is to provide a culture system and a method for culturing cells that can culture cells in parallel.

Means for Solving the Problems

[0005] The culture system according to an aspect of the present invention includes a cell container configured to contain cells, a plurality of culture devices, and a distribution flow path configured to distribute cells from the cell container to the plurality of culture devices. Each of the plurality of culture devices includes a plurality of culture tanks, a supply flow path configured to selectively supply fluid from the distribution flow path to the plurality of culture tanks, and a connection flow path configured to connect the plurality of culture tanks. In each of the plurality of culture devices, the plurality of culture tanks are connected in series by the connection flow path, and in each of the plurality of culture devices, the supply flow path is configured to selectively supply fluid to the plurality of culture tanks such that cells sequentially move through the plurality of culture tanks via the connection flow path.

[0006] The culture system described above may further include a fluid machine configured to supply fluid from a distribution channel to the supply channel of each of the multiple culture devices.

[0007] In the culture system described above, each of the multiple culture devices may further be provided with multiple supply valves located in the supply channel.

[0008] In the culture system described above, each of the multiple culture devices may have multiple supply valves, each corresponding to a multiple culture tank.

[0009] In the culture system described above, each of the multiple culture devices may further be provided with a connecting valve in the connecting channel.

[0010] In the culture system described above, connecting valves may be provided between multiple culture tanks.

[0011] In the culture system described above, each of the multiple culture devices may further include an intermediate tank provided in the connecting channel.

[0012] In the culture system described above, an intermediate tank may be provided between multiple culture tanks.

[0013] In the culture system described above, each of the multiple culture devices may further include a discharge channel configured to selectively discharge fluid from the multiple culture tanks.

[0014] In the culture system described above, each of the multiple culture devices may further be provided with multiple discharge valves in the discharge channel.

[0015] In the culture system described above, each of the multiple culture devices may have multiple discharge valves, each corresponding to a multiple culture tank.

[0016] In the culture system described above, each of the multiple culture devices may be configured such that a discharge channel selectively discharges fluid from multiple culture tanks so that cells move sequentially through multiple culture tanks via a connecting channel.

[0017] In the culture system described above, each of the multiple culture devices further comprises a plurality of supply valves provided in a supply channel, a discharge channel configured to selectively discharge fluid from a plurality of culture tanks, and a plurality of discharge valves provided in the discharge channel, wherein in each of the multiple culture devices, the plurality of supply valves each correspond to a plurality of culture tanks, and in each of the multiple culture devices, the plurality of discharge valves each correspond to a plurality of culture tanks, and one of the supply valves corresponding to one of the culture tanks from which cells are supplied from the cell container may be opened, and one of the discharge valves corresponding to one of the culture tanks from which cells are supplied from the cell container may be opened.

[0018] In the culture system described above, each of the multiple culture devices further comprises a plurality of supply valves provided in a supply channel, a discharge channel configured to selectively discharge fluid from a plurality of culture tanks, and a plurality of discharge valves provided in the discharge channel, wherein in each of the multiple culture devices, the plurality of supply valves each correspond to a plurality of culture tanks, and in each of the multiple culture devices, the plurality of discharge valves each correspond to a plurality of culture tanks, and one of the supply valves corresponding to one of the culture tanks to which fluid is supplied may open, and one of the discharge valves corresponding to one of the culture tanks to which fluid is supplied may open.

[0019] In the culture system described above, each of the multiple culture devices further comprises a plurality of supply valves provided in a supply channel, a plurality of connecting valves provided in a connecting channel, a discharge channel configured to selectively discharge fluid from a plurality of culture tanks, and a plurality of discharge valves provided in the discharge channel, wherein in each of the multiple culture devices, the plurality of supply valves each correspond to a plurality of culture tanks, and in each of the multiple culture devices, the plurality of connecting valves are provided between the plurality of culture tanks, and in each of the multiple culture devices, the plurality of discharge valves each correspond to a plurality of culture tanks, and one of the plurality of supply valves corresponding to one of the culture tanks in which cells are placed may be opened, the connecting valve between the culture tank in which cells are placed and the culture tank adjacent to the culture tank in which cells are placed may be opened, and one of the plurality of discharge valves corresponding to the adjacent culture tank may be opened.

[0020] In the culture system described above, one of the multiple discharge valves corresponding to the culture tank in which the cells are placed may be closed, and one of the multiple supply valves corresponding to the adjacent culture tank may be closed.

[0021] The culture system described above may further include one or more fluid containers configured to contain a fluid, and a distribution channel may be configured to distribute the fluid from the fluid container to a plurality of culture devices.

[0022] In the culture system described above, the fluid may contain a culture medium, a buffer, a cell release agent, a cryopreservation agent, or a factor.

[0023] The cell culture method according to an aspect of the present invention is a culture system including a cell container configured to accommodate cells, a plurality of culture devices, and a distribution flow path configured to distribute cells from the cell container to the plurality of culture devices. Each of the plurality of culture devices includes a plurality of culture tanks, a supply flow path configured to selectively supply fluid from the distribution flow path to the plurality of culture tanks, and a connection flow path configured to connect the plurality of culture tanks. The method includes preparing the culture system, distributing cells from the cell container to each of the plurality of culture devices via the distribution flow path, supplying cells to at least one of the plurality of culture tanks in each of the plurality of culture devices via the supply flow path, and culturing cells in at least one of the plurality of culture tanks in each of the plurality of culture devices.

[0024] In the above cell culture method, each of the plurality of culture devices further includes a discharge flow path configured to selectively discharge fluid from the plurality of culture tanks. When supplying cells to at least one of the plurality of culture tanks in each of the plurality of culture devices via the supply flow path, the fluid in at least one of the plurality of culture tanks may be discharged to the discharge flow path.

[0025] The above cell culture method may further include supplying fluid to the culture tank in which cells are placed among the plurality of culture tanks in each of the plurality of culture devices via the supply flow path.

[0026] In the above cell culture method, each of the plurality of culture devices further includes a discharge flow path configured to selectively discharge fluid from the plurality of culture tanks. When supplying fluid to the culture tank in which cells are placed among the plurality of culture tanks in each of the plurality of culture devices via the supply flow path, the fluid in the culture tank in which cells are placed among the plurality of culture tanks may be discharged to the discharge flow path.

[0027] The above-described method for culturing cells may further include moving cells from one culture vessel containing cells to another culture vessel via a connecting channel in each of the multiple culture devices.

[0028] In the cell culture method described above, each of the multiple culture devices is further provided with a discharge channel configured to selectively discharge fluid from the multiple culture tanks, and in each of the multiple culture devices, when moving cells from one culture tank to another via a connecting channel, the fluid in the other culture tank may be discharged into the discharge channel.

[0029] In the cell culture method described above, the fluid may be a gas, a liquid, or a gel.

[0030] In the cell culture method described above, the fluid may contain a culture medium, a buffer, a cell release agent, a cryopreservation agent, or a factor.

[0031] In the cell culture method described above, the fluid may contain a factor, and in each of the multiple culture devices, the factor may induce cells in different states from the cells in at least one of the multiple culture tanks. [Effects of the Invention]

[0032] According to the present invention, it is possible to provide a culture system and a cell culture method that enable the simultaneous cultivation of cells. [Brief explanation of the drawing]

[0033] [Figure 1] This is a schematic diagram of the culture system according to the first embodiment. [Figure 2] This is a schematic diagram of the culture system according to the first embodiment. [Figure 3] This is a schematic diagram of the culture system according to the second embodiment. [Figure 4] This is a schematic diagram of the culture system according to the third embodiment. [Figure 5] This is a schematic diagram of the culture system according to the fourth embodiment. [Modes for carrying out the invention]

[0034] Embodiments of the present invention are described below. In the following drawings, identical or similar parts are denoted by identical or similar reference numerals. However, the drawings are schematic. Therefore, specific dimensions and other details should be determined by referring to the following description. It should also be noted that there are parts in the drawings where the relationships and ratios of dimensions differ from one another.

[0035] (First Embodiment) The culture system according to the first embodiment comprises a cell container 10 configured to contain cells, a plurality of culture devices 20A, 20B, and 20C, and a distribution channel 30 configured to distribute cells from the cell container 10 to the plurality of culture devices 20A, 20B, and 20C. The number of the plurality of culture devices 20A, 20B, and 20C is not particularly limited.

[0036] Each of the multiple culture devices 20A, 20B, and 20C comprises multiple culture tanks 21, a supply channel 22 configured to selectively supply fluid from the distribution channel 30 to the multiple culture tanks 21, and a connecting channel 23 connecting the multiple culture tanks 21. The number of multiple culture tanks 21 is not particularly limited. Here, fluid includes gas, liquid, and gel.

[0037] In each of the multiple culture devices 20A, 20B, and 20C, the multiple culture tanks 21 are connected in series by a connecting channel 23. In addition, in each of the multiple culture devices 20A, 20B, and 20C, the supply channel 22 is configured to selectively supply fluid to the multiple culture tanks 21 so that cells move sequentially through the multiple culture tanks 21 via the connecting channel 23.

[0038] The culture system may further include one or more fluid containers 11A, 11B, 11C, and 11D. The number of fluid containers 11A, 11B, 11C, and 11D is not particularly limited. The cells contained in the cell container 10 are not particularly limited. The fluids contained in the fluid containers 11A, 11B, 11C, and 11D are not particularly limited. For example, the first fluid container 11A contains culture medium. The second fluid container 11B contains a release agent. The third fluid container 11C contains a cryopreservative. The fourth fluid container 11D contains gas. For example, the volumes of the cell container 10 and the fluid containers 11A, 11B, 11C, and 11D are variable.

[0039] For example, the cell container 10 and the fluid containers 11A, 11B, 11C, and 11D are connected to the confluence channel 32. The cell container 10 and the fluid containers 11A, 11B, 11C, and 11D may be connected aseptically to the confluence channel 32. The confluence channel 32 is connected to the distribution channel 30. The confluence channel 32 is provided with multiple confluence valves 33, 34A, 34B, 34C, and 34D, corresponding to the cell container 10 and the fluid containers 11A, 11B, 11C, and 11D, respectively. Note that if ambient air is used as the gas, the fluid container 11D does not need to be connected to the confluence channel 32. In this case, ambient air can be introduced into the confluence channel 32 by opening the confluence valve 34D. A filter may be provided at the opening of the confluence channel 32 into which ambient air is introduced.

[0040] When cells are transferred from the cell container 10 to the distribution channel 30 via the confluence channel 32, the confluence valve 33 corresponding to the cell container 10 is opened, and the other confluence valves 34A, 34B, 34C, and 34D are closed.

[0041] When the culture medium is sent from the first fluid container 11A to the distribution channel 30 via the confluence channel 32, the confluence valve 34A corresponding to the first fluid container 11A is opened, and the other confluence valves 33, 34B, 34C, and 34D are closed.

[0042] When the release agent is sent from the second fluid container 11B to the distribution channel 30 via the confluence channel 32, the confluence valve 34B corresponding to the second fluid container 11B is opened, and the other confluence valves 33, 34A, 34C, and 34D are closed.

[0043] When the cryopreservative is delivered from the third fluid container 11C to the distribution channel 30 via the confluence channel 32, the confluence valve 34C corresponding to the third fluid container 11C is opened, and the other confluence valves 33, 34A, 34B, and 34D are closed.

[0044] When gas is supplied from the fourth fluid container 11D to the distribution channel 30 via the confluence channel 32, the confluence valve 34D corresponding to the fourth fluid container 11D is opened, and the other confluence valves 33, 34A, 34B, and 34C are closed.

[0045] The distribution channel 30 connects the cell container 10 and one of the fluid containers 11A, 11B, 11C, or 11D to the multiple culture devices 20A, 20B, or 20C. The distribution channel 30 branches out and connects to the multiple culture devices 20A, 20B, or 20C. The distribution channel 30 may be equipped with a fluid machine 31 for moving the fluid within the distribution channel 30. The fluid machine 31 is, for example, a pump.

[0046] The cells in the cell container 10 are distributed to multiple culture devices 20A, 20B, and 20C by the fluid machine 31 via the confluence channel 32 and the distribution channel 30. For example, the type and amount of cells distributed to the multiple culture devices 20A, 20B, and 20C may be the same. Alternatively, at least one of the types and amounts of cells distributed to the multiple culture devices 20A, 20B, and 20C may be different.

[0047] The culture medium in the first fluid container 11A is distributed to multiple culture devices 20A, 20B, and 20C by the fluid machine 31 via the confluence channel 32 and the distribution channel 30. For example, the type and amount of culture medium distributed to the multiple culture devices 20A, 20B, and 20C may be the same. Alternatively, the amount of culture medium distributed to the multiple culture devices 20A, 20B, and 20C may be different.

[0048] The release agent in the second fluid container 11B is distributed to the multiple culture devices 20A, 20B, and 20C by the fluid machine 31 via the confluence channel 32 and the distribution channel 30. For example, the type and amount of release agent distributed to the multiple culture devices 20A, 20B, and 20C may be the same. Alternatively, the amount of release agent distributed to the multiple culture devices 20A, 20B, and 20C may be different.

[0049] The cryopreservative in the third fluid container 11C is distributed to multiple culture devices 20A, 20B, and 20C by the fluid machine 31 via the confluence channel 32 and the distribution channel 30. For example, the type and amount of cryopreservative distributed to the multiple culture devices 20A, 20B, and 20C may be the same. Alternatively, the amount of cryopreservative distributed to the multiple culture devices 20A, 20B, and 20C may be different.

[0050] The gas in the fourth fluid container 11D is distributed to the multiple culture devices 20A, 20B, and 20C by the fluid machine 31 via the confluence channel 32 and the distribution channel 30. For example, the type and amount of gas distributed to the multiple culture devices 20A, 20B, and 20C may be the same. Alternatively, the amount of gas distributed to the multiple culture devices 20A, 20B, and 20C may be different.

[0051] In each of the multiple culture devices 20A, 20B, and 20C, a supply channel 22 is connected to a distribution channel 30. The supply channel 22 branches out and connects to multiple culture tanks 21. Each branched supply channel 22 is provided with multiple supply valves 24 corresponding to the multiple culture tanks 21. When supplying fluid to any one of the multiple culture tanks 21, one corresponding supply valve 24 is opened and the other supply valves 24 are closed.

[0052] Each of the multiple culture devices 20A, 20B, and 20C may further include one or more connecting valves 25 provided in the connecting channel 23. For example, each of the multiple connecting valves 25 is positioned between adjacent culture tanks 21. When fluid is moved between any pair of culture tanks 21, the connecting valve 25 between that pair of culture tanks 21 is opened and the other connecting valves 25 are closed.

[0053] Each of the multiple culture devices 20A, 20B, and 20C may further be provided with a discharge channel 26 for selectively discharging fluid from multiple culture tanks 21. The discharge channel 26 is connected to the multiple culture tanks 21. The discharge channel 26 is provided with multiple discharge valves 27, each corresponding to one of the multiple culture tanks 21. When fluid is discharged from any one of the multiple culture tanks 21, one corresponding discharge valve 27 is opened and the other discharge valves 27 are closed. As shown in Figure 2, the supply channel 22 may be connected to a higher position in the direction of gravity relative to the culture tank 21, while the discharge channel 26 and connecting channel 23 may be connected to a lower position.

[0054] A cell recovery container 41 may be connected to at least one of the culture tanks 21 in each of the multiple culture devices 20A, 20B, and 20C shown in Figure 1, via a cell recovery channel 40. A recovery valve 28 may be provided in the cell recovery channel 40. The cell recovery container 41 may be connected to the cell recovery channel 40 in a sterile manner. A discharge container 50 may be connected to the discharge channel 26 of each of the multiple culture devices 20A, 20B, and 20C. For example, the volumes of the cell recovery container 41 and the discharge container 50 are variable.

[0055] The culture system according to the first embodiment may further include a control unit 301 that controls the fluid machine 31, the confluence valves 33 and 34, the supply valve 24, the discharge valve 27, and the recovery valve 28. The control unit 301 controls, for example, the flow rate, flow velocity, and fluid pressure of the fluid moved by the fluid machine 31. The control unit 301 controls the opening and closing of the confluence valves 33 and 34, the supply valve 24, the discharge valve 27, and the recovery valve 28, respectively, and controls the flow rate, flow velocity, and fluid pressure of the fluid passing through the confluence valves 33 and 34, the supply valve 24, the discharge valve 27, and the recovery valve 28, respectively.

[0056] Next, a method for culturing cells using the culture system according to the first embodiment will be described.

[0057] The control unit 301 opens the first supply valve 24AA and the first discharge valve 27AA corresponding to the first culture tank 21AA of the first culture apparatus 20A, closes the other supply valves 24AB, 24AC and the other discharge valves 27AB, 27AC of the first culture apparatus 20A, closes the connecting valves 25AA, 25AB, and closes the recovery valve 28.

[0058] The control unit 301 opens the first supply valve 24BA and the first discharge valve 27BA corresponding to the first culture tank 21BA of the second culture apparatus 20B, closes the other supply valves 24BB, 24BC and the other discharge valves 27BB, 27BC of the second culture apparatus 20B, closes the connecting valves 25BA, 25BB, and closes the recovery valve 28B.

[0059] The control unit 301 opens the first supply valve 24CA and the first discharge valve 27CA corresponding to the first culture tank 21CA of the third culture apparatus 20C, closes the other supply valves 24CB and 24CC and the other discharge valves 27CB and 27CC of the third culture apparatus 20C, closes the connecting valves 25CA and 25CB, and closes the recovery valve 28C.

[0060] The control unit 301 opens the confluence valve 33 corresponding to the cell container 10 and closes the other confluence valves 34A, 34B, 34C, and 34D. The control unit 301 controls the fluid machine 31, which moves the fluid in the distribution channel 30 toward the first to third culture devices 20A, 20B, and 20C. This moves the fluid containing cells in the cell container 10 through the confluence channel 32 to the distribution channel 30. The fluid containing cells is, for example, a culture medium. Next, the control unit 301 opens the confluence valve 34D corresponding to the fourth fluid container 11D containing gas and closes the other confluence valves 33, 34A, 34B, and 34C. This moves the gas, following the fluid containing cells, through the confluence channel 32 to the distribution channel 30. The fluid containing cells is then distributed by the distribution channel 30 to the first to third culture devices 20A, 20B, and 20C. Following the fluid containing cells, a gas is distributed through the distribution channel 30 to the first to third culture devices 20A, 20B, and 20C.

[0061] The fluid containing cells, which has been moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the first culture tank 21AA. The fluid, such as gas, inside the first culture tank 21AA is pushed by the fluid containing cells and is moved through the discharge channel 26A to the discharge container 50.

[0062] The fluid containing cells, which has been moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the first culture tank 21BA. The fluid, such as gas, inside the first culture tank 21BA is pushed by the fluid containing cells and is moved through the discharge channel 26B to the discharge container 50.

[0063] The fluid containing cells, which has been moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the first culture tank 21CA. The fluid, such as gas, inside the first culture tank 21CA is pushed by the fluid containing cells and is moved through the discharge channel 26C to the discharge container 50.

[0064] The length of the distribution channel 30 and / or supply channel 22 may be set so that the fluid containing cells reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the cell container 10 to each of the first culture tanks 21AA, 21BA, and 21CA are the same. Alternatively, the timing of the opening of the first supply valves 24AA, 24BA, and 24CA may be made different so that the fluid containing cells reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C at different times.

[0065] When the fluid containing cells enters the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, which stops moving the fluid. The control unit 301 closes the first supply valve 24AA and the first discharge valve 27AA of the first culture device 20A, the first supply valve 24BA and the first discharge valve 27BA of the second culture device 20B, and the first supply valve 24CA and the first discharge valve 27CA of the third culture device 20C. For a predetermined period, cells are cultured in each of the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C. The culture may be adherent culture or suspension culture. The culture may be two-dimensional culture or three-dimensional culture.

[0066] When changing the culture medium in the first culture tanks 21AA, 21BA, and 21CA, the control unit 301 opens the first supply valve 24AA and the first discharge valve 27AA of the first culture device 20A, and closes the other supply valves 24AB, 24AC and the other discharge valves 27AB, 27AC. The control unit 301 opens the first supply valve 24BA and the first discharge valve 27BA of the second culture device 20B, and closes the other supply valves 24BB, 24BC and the other discharge valves 27BB, 27BC. The control unit 301 opens the first supply valve 24CA and the first discharge valve 27CA of the third culture device 20C, and closes the other supply valves 24CB, 24CC and the other discharge valves 27CB, 27CC. Furthermore, the control unit 301 opens the confluence valve 34A and closes the other confluence valves 33, 34B, 34C, and 34D, and controls the fluid machine 31, which moves the unused culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0067] Unused culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the first culture tank 21AA. Used culture medium in the first culture tank 21AA is pushed by the unused culture medium and moved through the discharge channel 26A to the discharge container 50. This replaces the culture medium in the first culture tank 21AA.

[0068] Unused culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the first culture tank 21BA. Used culture medium in the first culture tank 21BA is pushed by the unused culture medium and moved through the discharge channel 26B to the discharge container 50. This replaces the culture medium in the first culture tank 21BA.

[0069] Unused culture medium, moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the first culture tank 21CA. Used culture medium in the first culture tank 21CA is pushed by the unused culture medium and moved through the discharge channel 26C to the discharge container 50. This replaces the culture medium in the first culture tank 21CA.

[0070] The length of the distribution channel 30 and / or supply channel 22 may be set so that the culture medium reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the first fluid container 11A to each of the first culture tanks 21AA, 21BA, and 21CA are the same. Alternatively, the timing of the opening of the first supply valves 24AA, 24BA, and 24CA may be made different so that the culture medium reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C at different times.

[0071] Before sending unused culture medium to each of the first culture tanks 21AA, 21BA, and 21CA, some or all of the culture medium in each of the first culture tanks 21AA, 21BA, and 21CA may be drained.

[0072] In this case, the control unit 301 opens the confluence valve 34D, closes the other confluence valves 33, 34A, 34B, and 34C, and controls the fluid machine 31, which moves the gas in the fourth fluid container 11D towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0073] The gas moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A reaches the first culture tank 21AA. Some or all of the used culture medium in the first culture tank 21AA is pushed by the gas and moved through the discharge channel 26A to the discharge container 50. As a result, some or all of the culture medium in the first culture tank 21AA is discharged.

[0074] The gas moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B reaches the first culture tank 21BA. Some or all of the used culture medium in the first culture tank 21BA is pushed by the gas and moved through the discharge channel 26B to the discharge container 50. As a result, some or all of the culture medium in the first culture tank 21BA is discharged.

[0075] The gas moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C reaches the first culture tank 21CA. Some or all of the used culture medium in the first culture tank 21CA is pushed by the gas and moved through the discharge channel 26C to the discharge container 50. As a result, some or all of the culture medium in the first culture tank 21CA is discharged.

[0076] When the exchange of culture medium in the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C is completed, the control unit 301 controls the fluid machine 31, which stops moving the fluid. The control unit 301 closes the first supply valve 24AA and the first discharge valve 27AA of the first culture device 20A, the first supply valve 24BA and the first discharge valve 27BA of the second culture device 20B, and the first supply valve 24CA and the first discharge valve 27CA of the third culture device 20C. Cells are cultured in each of the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C for a predetermined period. The culture may be adherent culture or suspension culture. The culture may be two-dimensional culture or three-dimensional culture.

[0077] When subculturing cells in the first culture tanks 21AA, 21BA, and 21CA, the control unit 301 opens the first supply valve 24AA and the first discharge valve 27AA of the first culture device 20A, and closes the other supply valves 24AB, 24AC and the other discharge valves 27AB, 27AC. The control unit 301 opens the first supply valve 24BA and the first discharge valve 27BA of the second culture device 20B, and closes the other supply valves 24BB, 24BC and the other discharge valves 27BB, 27BC. The control unit 301 opens the first supply valve 24CA and the first discharge valve 27CA of the third culture device 20C, and closes the other supply valves 24CB, 24CC and the other discharge valves 27CB, 27CC. Furthermore, the control unit 301 opens the confluence valve 34B and closes the other confluence valves 33, 34A, 34C, and 34D, and controls the fluid machine 31, which moves the release agent in the second fluid container 11B towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0078] The release agent, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the first culture tank 21AA. The used culture medium in the first culture tank 21AA is pushed by the release agent and moved through the discharge channel 26A to the discharge container 50.

[0079] The release agent, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the first culture tank 21BA. The used culture medium in the first culture tank 21BA is pushed by the release agent and moved through the discharge channel 26B to the discharge container 50.

[0080] The release agent, moved from the distribution channel 30 to the supply channel 22C of the third culture device 20C, reaches the first culture tank 21CA. The used culture medium in the first culture tank 21CA is pushed by the release agent and moved through the discharge channel 26C to the discharge container 50.

[0081] The length of the distribution channel 30 and / or supply channel 22 may be set so that the release agent reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the second fluid container 11B to each of the first culture tanks 21AA, 21BA, and 21CA are the same. Alternatively, the timing of the opening of the first supply valves 24AA, 24BA, and 24CA may be made different so that the release agent reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C at different times.

[0082] Furthermore, before sending the release agent to each of the first culture tanks 21AA, 21BA, and 21CA, some or all of the culture medium in each of the first culture tanks 21AA, 21BA, and 21CA may be discharged using gas. The method for discharging some or all of the culture medium in each of the first culture tanks 21AA, 21BA, and 21CA using gas is as described above.

[0083] When the detachment agent is introduced into the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, which stops moving the fluid. The cells are then brought into contact with the detachment agent for a predetermined time to detach them from each of the first culture tanks 21AA, 21BA, and 21CA. In the case of suspension culture or three-dimensional culture, it is not necessary to bring the cells into contact with the detachment agent.

[0084] After a predetermined time has elapsed, the control unit 301 closes the confluence valves 33, 34A, 34B, and 34C, and opens the confluence valve 34D. The control unit 301 also controls the fluid machine 31, which moves the gas in the fourth fluid container 11D towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0085] The gas moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A reaches the first culture tank 21AA. The used release agent in the first culture tank 21AA is pushed by the gas and moved through the discharge channel 26A to the discharge container 50.

[0086] The gas moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B reaches the first culture tank 21BA. The used release agent in the first culture tank 21BA is pushed by the gas and moved through the discharge channel 26B to the discharge container 50.

[0087] The gas moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C reaches the first culture tank 21CA. The used release agent in the first culture tank 21CA is pushed by the gas and moved through the discharge channel 26C to the discharge container 50.

[0088] The length of the distribution channel 30 and / or supply channel 22 may be set so that the gas reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the flow path length from the fourth fluid container 11D to each of the first culture tanks 21AA, 21BA, and 21CA is the same. Alternatively, the opening timing of the first supply valves 24AA, 24BA, and 24CA may be made different so that the gas reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C at different times.

[0089] The control unit 301 opens the first supply valve 24AA corresponding to the first culture tank 21AA of the first culture apparatus 20A, closes the first discharge valve 27AA corresponding to the first culture tank 21AA, closes the second supply valve 24AB corresponding to the second culture tank 21AB, opens the second discharge valve 27AB corresponding to the second culture tank 21AB, closes the third supply valve 24AC corresponding to the third culture tank 21AC, and closes the third discharge valve 27AC corresponding to the third culture tank 21AC. The control unit 301 opens the first connecting valve 25AA between the first culture tank 21AA and the second culture tank 21AB of the first culture apparatus 20A, and closes the second connecting valve 25AB between the second culture tank 21AB and the third culture tank 21AC.

[0090] The control unit 301 opens the first supply valve 24BA corresponding to the first culture tank 21BA of the second culture apparatus 20B, closes the first discharge valve 27BA corresponding to the first culture tank 21BA, closes the second supply valve 24BB corresponding to the second culture tank 21BB, opens the second discharge valve 27BB corresponding to the second culture tank 21BB, closes the third supply valve 24BC corresponding to the third culture tank 21BC, and closes the third discharge valve 27BC corresponding to the third culture tank 21BC. The control unit 301 opens the first connecting valve 25BA between the first culture tank 21BA and the second culture tank 21BB of the second culture apparatus 20B, and closes the second connecting valve 25BB between the second culture tank 21BB and the third culture tank 21BC.

[0091] The control unit 301 opens the first supply valve 24CA corresponding to the first culture tank 21CA of the third culture apparatus 20C, closes the first discharge valve 27CA corresponding to the first culture tank 21CA, closes the second supply valve 24CB corresponding to the second culture tank 21CB, opens the second discharge valve 27CB corresponding to the second culture tank 21CB, closes the third supply valve 24CC corresponding to the third culture tank 21CC, and closes the third discharge valve 27CC corresponding to the third culture tank 21CC. The control unit 301 opens the first connecting valve 25CA between the first culture tank 21CA and the second culture tank 21CB of the third culture apparatus 20C, and closes the second connecting valve 25CB between the second culture tank 21CB and the third culture tank 21CC.

[0092] The control unit 301 closes the confluence valves 33, 34B, 34C, and 34D, and opens the confluence valve 34A corresponding to the first fluid container 11A that contains the culture medium. The control unit 301 controls the fluid machine 31, which moves the culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0093] The culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the first culture tank 21AA. The cells in the first culture tank 21AA are dispersed in the culture medium. The culture medium containing the cells is moved to the second culture tank 21AB via the connecting channel 23A. The fluid, such as gas, in the second culture tank 21AB is pushed by the culture medium containing the cells and moved to the discharge container 50 via the discharge channel 26A.

[0094] Distribution channel 30 to the 2 The culture medium, moved through the supply channel 22B of the culture apparatus 20B, reaches the first culture tank 21BA. The cells in the first culture tank 21BA are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23B into the second culture tank 21BB. The fluid, such as gas, in the second culture tank 21BB is pushed by the culture medium containing the cells and is moved through the discharge channel 26B to the discharge container 50.

[0095] Distribution channel 30 to the 3The culture medium, moved through the supply channel 22B of the culture apparatus 20C, reaches the first culture tank 21CA. The cells in the first culture tank 21CA are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23C into the second culture tank 21CB. The fluid, such as gas, in the second culture tank 21CB is pushed by the culture medium containing the cells and moved through the discharge channel 26C to the discharge container 50.

[0096] The lengths of the connecting channels 23A, 23B, and 23C may be set so that the culture medium containing cells reaches the second culture tanks 21AB, 21BB, and 21CB from the first culture tanks 21AA, 21BA, and 21CA simultaneously. For example, the lengths of the connecting channels 23A, 23B, and 23C may be set so that the length of each channel from the first culture tanks 21AA, 21BA, and 21CA to the second culture tanks 21AB, 21BB, and 21CB is the same. Alternatively, the timing of the opening of the first supply valves 24AA, 24BA, and 24CA may be made different so that the culture medium reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C at different times, and the culture medium containing cells reaches the second culture tanks 21AB, 21BB, and 21CB from the first culture tanks 21AA, 21BA, and 21CA at different times.

[0097] When the fluid containing cells enters the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31 and stops the fluid machine 31 from moving the fluid. The control unit 301 closes the first supply valve 24AA, the first connecting valve 25AA, and the second discharge valve 27AB of the first culture device 20A, the first supply valve 24BA, the first connecting valve 25BA, and the second discharge valve 27BB of the second culture device 20B, and the first supply valve 24CA, the first connecting valve 25CA, and the second discharge valve 27CB of the third culture device 20C. Cells are cultured in each of the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C for a predetermined period of time. The culture may be adherent culture or suspension culture. The culture may be two-dimensional culture or three-dimensional culture.

[0098] When changing the culture medium in the second culture tanks 21AB, 21BB, and 21CB, the control unit 301 opens the second supply valve 24AB and the second discharge valve 27AB of the first culture device 20A, and closes the other supply valves 24AA, 24AC and the other discharge valves 27AA, 27AC. The control unit 301 opens the second supply valve 24BB and the second discharge valve 27BB of the second culture device 20B, and closes the other supply valves 24BA, 24BC and the other discharge valves 27BA, 27BC. The control unit 301 opens the second supply valve 24CB and the second discharge valve 27CB of the third culture device 20C, and closes the other supply valves 24CA, 24CC and the other discharge valves 27CA, 27CC. Furthermore, the control unit 301 opens the confluence valve 34A and closes the other confluence valves 33, 34B, 34C, and 34D, and controls the fluid machine 31, which moves the unused culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0099] Unused culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the second culture tank 21AB. The used culture medium in the second culture tank 21AB is pushed by the unused culture medium and moved through the discharge channel 26A to the discharge container 50. This replaces the culture medium in the second culture tank 21AB.

[0100] Unused culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the second culture tank 21BB. Used culture medium in the second culture tank 21BB is pushed by the unused culture medium and moved through the discharge channel 26B to the discharge container 50. This replaces the culture medium in the second culture tank 21BB.

[0101] Unused culture medium, moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the second culture tank 21CB. Used culture medium in the second culture tank 21CB is pushed by the unused culture medium and moved through the discharge channel 26C to the discharge container 50. This replaces the culture medium in the second culture tank 21CB.

[0102] The length of the distribution channel 30 and / or supply channel 22 may be set so that the culture medium reaches the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the first fluid container 11A to each of the second culture tanks 21AB, 21BB, and 21CB are the same. 2 The timing of the opening of the supply valves 24AB, 24BB, and 24CB is different, so that the culture medium is supplied to the first to third culture devices 20A, 20B, and 20C. 2 The cultures may be arranged to reach culture tanks 21AB, 21BB, and 21CB at different times.

[0103] When the exchange of culture medium in the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C is completed, the control unit 301 controls the fluid machine 31 and stops the fluid machine 31 from moving the fluid. The control unit 301 closes the second supply valve 24AB and the second discharge valve 27AB of the first culture device 20A, the second supply valve 24BB and the second discharge valve 27BB of the second culture device 20B, and the second supply valve 24CB and the second discharge valve 27CB of the third culture device 20C. Cells are cultured in each of the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C for a predetermined period. The culture may be adherent culture or suspension culture. The culture may be two-dimensional culture or three-dimensional culture.

[0104] When subculturing cells in the second culture tanks 21AB, 21BB, and 21CB, the control unit 301 opens the second supply valve 24AB and the second discharge valve 27AB of the first culture apparatus 20A, and closes the other supply valves 24AA, 24AC and the other discharge valves 27AA, 27AC. The control unit 301 opens the second supply valve 24BB and the second discharge valve 27BB of the second culture apparatus 20B, and closes the other supply valves 24BA, 24BC and the other discharge valves 27BA, 27BC. The control unit 301 opens the second supply valve 24CB and the second discharge valve 27CB of the third culture apparatus 20C, and closes the other supply valves 24CA, 24CC and the other discharge valves 27CA, 27CC. Furthermore, the control unit 301 opens the confluence valve 34B and closes the other confluence valves 33, 34A, 34C, and 34D, and controls the fluid machine 31, which moves the release agent in the second fluid container 11B towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0105] The release agent, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the second culture tank 21AB. The used culture medium in the second culture tank 21AB is pushed by the release agent and moved through the discharge channel 26A to the discharge container 50.

[0106] The release agent, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the second culture tank 21BB. The used culture medium in the second culture tank 21BB is pushed by the release agent and moved through the discharge channel 26B to the discharge container 50.

[0107] The release agent, moved from the distribution channel 30 to the supply channel 22C of the third culture device 20C, reaches the second culture tank 21CB. The used culture medium in the second culture tank 21CB is pushed by the release agent and moved through the discharge channel 26C to the discharge container 50.

[0108] The length of the distribution channel 30 and / or supply channel 22 may be set so that the release agent reaches the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the second fluid container 11B to each of the second culture tanks 21AB, 21BB, and 21CB are the same. 2 The supply valves 24AB, 24BB, and 24CB open at different times, allowing the release agent to be released into the first to third culture devices 20A, 20B, and 20C. 2 The cultures may be arranged to reach culture tanks 21AB, 21BB, and 21CB at different times.

[0109] When the detachment agent enters the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, causing the fluid machine 31 to stop moving the fluid. The cells are then brought into contact with the detachment agent for a predetermined time to detach them from each of the second culture tanks 21AB, 21BB, and 21CB. In the case of suspension culture or three-dimensional culture, it is not necessary to bring the cells into contact with the detachment agent.

[0110] After a predetermined time has elapsed, the control unit 301 closes the confluence valves 33, 34A, 34B, and 34C, and opens the confluence valve 34D. The control unit 301 also controls the fluid machine 31, which moves the gas in the fourth fluid container 11D towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0111] The gas, which has been moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the second culture tank 21AB. The used release agent in the second culture tank 21AB is pushed by the gas and moved through the discharge channel 26A to the discharge container 50.

[0112] The gas moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B reaches the second culture tank 21BB. The used release agent in the second culture tank 21BB is pushed by the gas and moved through the discharge channel 26B to the discharge container 50.

[0113] The gas moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C reaches the second culture tank 21CB. The used release agent in the second culture tank 21CB is pushed by the gas and moved through the discharge channel 26C to the discharge container 50.

[0114] The length of the distribution channel 30 and / or supply channel 22 may be set so that the gas reaches the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the flow path length from the fourth fluid container 11D to each of the second culture tanks 21AB, 21BB, and 21CB is the same. 2 The supply valves 24AB, 24BB, and 24CB open at different times, allowing gas to flow into the first to third culture devices 20A, 20B, and 20C. 2 The cultures may be arranged to reach culture tanks 21AB, 21BB, and 21CB at different times.

[0115] The control unit 301 closes the first supply valve 24AA and the first discharge valve 27AA of the first culture apparatus 20A, opens the second supply valve 24AB, closes the second discharge valve 27AB, closes the third supply valve 24AC, and opens the third discharge valve 27AC. The control unit 301 closes the first connecting valve 25AA and opens the second connecting valve 25AB of the first culture apparatus 20A. The control unit 301 closes the recovery valve 28A provided in the cell recovery channel 40A connected to the third culture tank 21AC.

[0116] The control unit 301 closes the first supply valve 24BA and the first discharge valve 27BA of the second culture apparatus 20B, opens the second supply valve 24BB, closes the second discharge valve 27BB, closes the third supply valve 24BC, and opens the third discharge valve 27BC. The control unit 301 closes the first connecting valve 25BA and opens the second connecting valve 25BB of the second culture apparatus 20B. The control unit 301 closes the recovery valve 28B provided in the cell recovery channel 40B connected to the third culture tank 21BC.

[0117] The control unit 301 closes the first supply valve 24CA and the first discharge valve 27CA of the third culture apparatus 20C, opens the second supply valve 24CB, closes the second discharge valve 27CB, closes the third supply valve 24CC, and opens the third discharge valve 27CC. The control unit 301 closes the first connecting valve 25CA and opens the second connecting valve 25CB of the third culture apparatus 20C. The control unit 301 closes the recovery valve 28C provided in the cell recovery channel 40C connected to the third culture tank 21CC.

[0118] The control unit 301 closes the confluence valves 33, 34B, 34C, and 34D, and opens the confluence valve 34A corresponding to the first fluid container 11A that contains the culture medium. The control unit 301 controls the fluid machine 31, which moves the culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0119] The culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the second culture tank 21AB. The cells in the second culture tank 21AB are dispersed in the culture medium. The culture medium containing the cells is moved to the third culture tank 21AC via the connecting channel 23A. The fluid, such as gas, in the third culture tank 21AC is pushed by the culture medium containing the cells and moved to the discharge container 50 via the discharge channel 26A.

[0120] The culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the second culture tank 21BB. The cells in the second culture tank 21BB are dispersed in the culture medium. The culture medium containing the cells is moved to the third culture tank 21BC via the connecting channel 23B. The fluid, such as gas, in the third culture tank 21BC is pushed by the culture medium containing the cells and moved to the discharge container 50 via the discharge channel 26B.

[0121] From the distribution channel 30 to the supply channel 22 of the third culture device 20C C The culture medium, which has been moved, reaches the second culture tank 21CB. The cells in the second culture tank 21CB are dispersed in the culture medium. The culture medium containing the cells is moved into the third culture tank 21CC via the connecting channel 23C. The fluid such as gas in the third culture tank 21CC is pushed by the culture medium containing the cells and is moved to the discharge container 50 via the discharge channel 26C.

[0122] The lengths of the connecting channels 23A, 23B, and 23C may be set so that the culture medium containing cells reaches the third culture tanks 21AC, 21BC, and 21CC simultaneously from the second culture tanks 21AB, 21BB, and 21CB, respectively. For example, the lengths of the connecting channels 23A, 23B, and 23C may be set so that the length of each channel from the second culture tanks 21AB, 21BB, and 21CB to the third culture tanks 21AC, 21BC, and 21CC is the same. The timing at which the first supply valves 24AB, 24BB, and 24CB open may be made different so that the culture medium reaches the first to third culture devices 20A, 20B, and 20C simultaneously. 2 The culture medium may reach the culture vessels 21AB, 21BB, and 21CB at different times, and the culture medium containing the cells may reach the third culture vessels 21AC, 21BC, and 21CC from the second culture vessels 21AB, 21BB, and 21CB at different times.

[0123] When the fluid containing cells enters the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31 and stops the fluid machine 31 from moving the fluid. The control unit 301 closes the second supply valve 24AB, the second connecting valve 25AB, and the third discharge valve 27AC of the first culture device 20A, the second supply valve 24BB, the second connecting valve 25BB, and the third discharge valve 27BC of the second culture device 20B, and the second supply valve 24CB, the second connecting valve 25CB, and the third discharge valve 27CC of the third culture device 20C. Cells are cultured in each of the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C for a predetermined period of time. The culture may be adherent culture or suspension culture. The culture may be two-dimensional culture or three-dimensional culture.

[0124] When changing the culture medium in the third culture tanks 21AC, 21BC, and 21CC, the control unit 301 opens the third supply valve 24AC and the third discharge valve 27AC of the first culture device 20A, and closes the other supply valves 24AA, 24AB and the other discharge valves 27AA, 27AB. The control unit 301 opens the third supply valve 24BC and the third discharge valve 27BC of the second culture device 20B, and closes the other supply valves 24BA, 24BB and the other discharge valves 27BA, 27BB. The control unit 301 opens the third supply valve 24CC and the third discharge valve 27CC of the third culture device 20C, and closes the other supply valves 24CA, 24CB and the other discharge valves 27CA, 27CB. Furthermore, the control unit 301 opens the confluence valve 34A and closes the other confluence valves 33, 34B, 34C, and 34D, and controls the fluid machine 31, which moves the unused culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0125] Unused culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the third culture tank 21AC. Used culture medium in the second culture tank 21AC is pushed by the unused culture medium and moved through the discharge channel 26A to the discharge container 50. This replaces the culture medium in the third culture tank 21AC.

[0126] Unused culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the third culture tank 21BC. The used culture medium in the third culture tank 21BC is pushed by the unused culture medium and moved through the discharge channel 26B to the discharge container 50. This replaces the culture medium in the third culture tank 21BC.

[0127] Unused culture medium, moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the third culture tank 21CC. Used culture medium in the third culture tank 21CC is pushed by the unused culture medium and moved through the discharge channel 26C to the discharge container 50. This replaces the culture medium in the third culture tank 21CC.

[0128] The length of the distribution channel 30 and / or supply channel 22 may be set so that the culture medium reaches the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the flow path length from the first fluid container 11A to each of the third culture tanks 21AC, 21BC, and 21CC is the same. 3 The timing of opening the supply valves 24AC, 24BC, and 24CC is different, so that the culture medium is supplied to the first to third culture devices 20A, 20B, and 20C. 3 The culture vessels 21AC, 21BC, and 21CC may be arranged to reach the cells at different times.

[0129] When the exchange of culture medium in the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C is completed, the control unit 301 controls the fluid machine 31 and stops the fluid machine 31 from moving the fluid. The control unit 301 closes the third supply valve 24AC and the third discharge valve 27AC of the first culture device 20A, the third supply valve 24BC and the third discharge valve 27BC of the second culture device 20B, and the third supply valve 24CC and the third discharge valve 27CC of the third culture device 20C. Cells are cultured in each of the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C for a predetermined period. The culture may be adherent culture or suspension culture. The culture may be two-dimensional culture or three-dimensional culture.

[0130] When harvesting cells from the third culture tanks 21AC, 21BC, and 21CC, the control unit 301 opens the third supply valve 24AC and the third discharge valve 27AC of the first culture device 20A, and closes the other supply valves 24AA, 24AB and the other discharge valves 27AA, 27AB. The control unit 301 opens the third supply valve 24BC and the third discharge valve 27BC of the second culture device 20B, and closes the other supply valves 24BA, 24BB and the other discharge valves 27BA, 27BB. The control unit 301 opens the third supply valve 24CC and the third discharge valve 27CC of the third culture device 20C, and closes the other supply valves 24CA, 24CB and the other discharge valves 27CA, 27CB. Furthermore, the control unit 301 opens the confluence valve 34B and closes the other confluence valves 33, 34A, 34C, and 34D, and controls the fluid machine 31, which moves the release agent in the second fluid container 11B towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0131] The release agent, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the third culture tank 21AC. The used culture medium in the third culture tank 21AC is pushed by the release agent and moved through the discharge channel 26A to the discharge container 50.

[0132] The release agent, moved from the distribution channel 30 to the supply channel 22B of the second culture device 20B, reaches the third culture tank 21BC. The used culture medium in the third culture tank 21BC is pushed by the release agent and moved through the discharge channel 26B to the discharge container 50.

[0133] The release agent, moved from the distribution channel 30 to the supply channel 22C of the third culture device 20C, reaches the third culture tank 21CC. The used culture medium in the third culture tank 21CC is pushed by the release agent and moved through the discharge channel 26C to the discharge container 50.

[0134] The length of the distribution channel 30 and / or supply channel 22 may be set so that the release agent reaches the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the flow path length from the second fluid container 11B to each of the third culture tanks 21AC, 21BC, and 21CC is the same. 3 The supply valves 24AC, 24BC, and 24CC open at different times, allowing the release agent to be released into the first to third culture devices 20A, 20B, and 20C. 3 The culture vessels 21AC, 21BC, and 21CC may be arranged to reach the cells at different times.

[0135] When the detachment agent enters the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, causing the fluid machine 31 to stop moving the fluid. The cells are then brought into contact with the detachment agent for a predetermined time to detach them from each of the third culture tanks 21AC, 21BC, and 21CC. In the case of suspension culture or three-dimensional culture, it is not necessary to bring the cells into contact with the detachment agent.

[0136] After a predetermined time has elapsed, the control unit 301 closes the confluence valves 33, 34A, 34B, and 34C, and opens the confluence valve 34D. The control unit 301 also controls the fluid machine 31, which moves the gas in the fourth fluid container 11D towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0137] The gas moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A reaches the third culture tank 21AC. The used release agent in the third culture tank 21AC is pushed by the gas and moved through the discharge channel 26A to the discharge container 50.

[0138] The gas moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B reaches the third culture tank 21BC. The used release agent in the third culture tank 21BC is pushed by the gas and moved through the discharge channel 26B to the discharge container 50.

[0139] The gas moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C reaches the third culture tank 21CC. The used release agent in the third culture tank 21CC is pushed by the gas and moved through the discharge channel 26C to the discharge container 50.

[0140] The length of the distribution channel 30 and / or supply channel 22 may be set so that the gas reaches the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the flow path length from the fourth fluid container 11D to each of the third culture tanks 21AC, 21BC, and 21CC is the same. 3 The supply valves 24AC, 24BC, and 24CC open at different times, allowing gas to flow into the first to third culture devices 20A, 20B, and 20C. 3 The culture vessels 21AC, 21BC, and 21CC may be arranged to reach the cells at different times.

[0141] The control unit 301 closes the first supply valve 24AA of the first culture apparatus 20A, closes the first discharge valve 27AA, closes the second supply valve 24AB, closes the second discharge valve 27AB, opens the third supply valve 24AC, and closes the third discharge valve 27AC. The control unit 301 closes the first connecting valve 25AA of the first culture apparatus 20A, and closes the second connecting valve 25AB. The control unit 301 opens the recovery valve 28A.

[0142] The control unit 301 closes the first supply valve 24BA of the second culture device 20B, closes the first discharge valve 27BA, closes the second supply valve 24BB, closes the second discharge valve 27BB, opens the third supply valve 24BC, and closes the third discharge valve 27BC. The control unit 301 closes the first connecting valve 25BA of the second culture device 20B and closes the second connecting valve 25BB. The control unit 301 opens the recovery valve 28B.

[0143] The control unit 301 closes the first supply valve 24CA, the first discharge valve 27CA, the second supply valve 24CB, the second discharge valve 27CB of the third culture device 20C, opens the third supply valve 24CC, and closes the third discharge valve 27CC. The control unit 301 closes the first connecting valve 25CA and the second connecting valve 25CB of the third culture device 20C. The control unit 301 opens the recovery valve 28C.

[0144] The control unit 301 closes the merging valves 33, 34A, 34B, and 34D, and opens the merging valve 34C corresponding to the third fluid container 11C that contains the cryopreservative. The control unit 301 controls the fluid machine 31, which moves the cryopreservative in the third fluid container 11C towards the first to third culture devices 20A, 20B, and 20C via the merging channel 32 and the distribution channel 30.

[0145] The cryopreserved agent, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the third culture tank 21AC. The cells in the third culture tank 21AC are dispersed in the cryopreserved agent. The cryopreserved agent containing the cells is moved through the cell recovery channel 40A into the cell recovery container 41A.

[0146] The cryopreserved agent, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the third culture tank 21BC. The cells in the third culture tank 21BC are dispersed in the cryopreserved agent. The cryopreserved agent containing the cells is moved through the cell recovery channel 40B into the cell recovery container 41B.

[0147] The cryopreserved agent, moved from the distribution channel 30 to the supply channel 22B of the third culture apparatus 20C, reaches the third culture vessel 21CC. The cells in the third culture vessel 21CC are dispersed in the cryopreserved agent. The cryopreserved agent containing the cells is moved through the cell recovery channel 40C into the cell recovery container 41C.

[0148] The length of the distribution channel 30 and / or supply channel 22 may be set so that the cryopreserving agent reaches the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the flow path length from the third fluid container 11C to each of the third culture tanks 21AC, 21BC, and 21CC is the same. The timing of opening of the first supply valves 24AC, 24BC, and 24CC may be made different so that the cryopreserving agent reaches the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C simultaneously. 3 The culture vessels 21AC, 21BC, and 21CC may be arranged to reach the cells at different times.

[0149] When the fluid containing cells enters the cell recovery containers 41A, 41B, and 41C, the control unit 301 controls the fluid machine 31, causing it to stop moving the fluid. The control unit 301 closes the recovery valves 28A, 28B, and 28C. After that, the cell recovery containers 41A, 41B, and 41C may be removed from the cell recovery channels 40A, 40B, and 40C and cryopreserved.

[0150] In addition, Cell collection containers 41A, 41B, 41C When preserving cells, fluids other than cryopreservatives may be used, and culture media or buffer solutions may also be used.

[0151] (Second Embodiment) In the culture system according to the second embodiment, as shown in Figure 3, each of the multiple culture devices 20A, 20B, and 20C further comprises an intermediate tank 60 provided in the connecting channel 23. The intermediate tank 60 is provided between the culture tanks 21. The intermediate tank 60 is configured to contain cells. The intermediate tank 60 may be equipped with a stirring device configured to agitate the fluid containing the cells. The intermediate tank 60 may be equipped with a cell aggregate separation mechanism capable of separating cell aggregates. Other components of the culture system according to the second embodiment are the same as in the first embodiment.

[0152] In the second embodiment, when moving cells from the first culture tanks 21AA, 21BA, and 21CA to the second culture tanks 21AB, 21BB, and 21CB, if the cells are in adherent culture, the cells are detached from the culture tank, as in the first embodiment. If the cells are not in adherent culture, it is not necessary to detach the cells from the culture tank. Subsequently, the control unit 301 controls the valve and the fluid machine 31, as in the first embodiment, and the fluid machine 31 moves the culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0153] The culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the first culture tank 21AA. The cells in the first culture tank 21AA are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23A into the first intermediate tank 60AA. The fluid, such as gas, in the first intermediate tank 60AA is pushed by the culture medium containing the cells and is moved through the connecting channel 23A, the second culture tank 21AB, and the discharge channel 26A to the discharge container 50.

[0154] The culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the first culture tank 21BA. The cells in the first culture tank 21BA are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23B into the first intermediate tank 60BA. The fluid, such as gas, in the first intermediate tank 60BA is pushed by the culture medium containing the cells and is moved through the connecting channel 23B, the second culture tank 21BB, and the discharge channel 26B to the discharge container 50.

[0155] The culture medium, moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the first culture tank 21CA. The cells in the first culture tank 21CA are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23C into the first intermediate tank 60CA. The fluid, such as gas, in the first intermediate tank 60CA is pushed by the culture medium containing the cells and is moved through the connecting channel 23C, the second culture tank 21CB, and the discharge channel 26C to the discharge container 50.

[0156] When culture medium containing cells is placed in the first intermediate tanks 60AA, 60BA, and 60CA of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, which stops moving the fluid.

[0157] For example, after the cell density in the culture medium becomes uniform in each of the first intermediate tanks 60AA, 60BA, and 60CA, the control unit 301 controls the fluid machine 31, which moves the culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0158] The culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the first intermediate tank 60AA. The culture medium containing cells in the first intermediate tank 60AA is moved through the connecting channel 23A into the second culture tank 21AB. The fluid such as gas in the second culture tank 21AB is pushed by the culture medium containing cells and moved through the discharge channel 26A to the discharge container 50.

[0159] The culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the first intermediate tank 60BA. The culture medium containing cells in the first intermediate tank 60BA is moved through the connecting channel 23B into the second culture tank 21BB. The fluid such as gas in the second culture tank 21BB is pushed by the culture medium containing cells and moved through the discharge channel 26B to the discharge container 50.

[0160] The culture medium, moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the first intermediate tank 60CA. The culture medium containing cells in the first intermediate tank 60CA is moved through the connecting channel 23C into the second culture tank 21CB. The fluid, such as gas, in the second culture tank 21CB is pushed by the culture medium containing cells and moved through the discharge channel 26C to the discharge container 50.

[0161] In the second embodiment, when moving cells from the second culture tanks 21AB, 21BB, and 21CB into the third culture tanks 21AC, 21BC, and 21CC, the control unit 301 controls the valves and the fluid machine 31 in the same manner as in the first embodiment, and the fluid machine 31 moves the culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0162] The culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the second culture tank 21AB. The cells in the second culture tank 21AB are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23A into the second intermediate tank 60AB. The fluid such as gas in the second intermediate tank 60AB is pushed by the culture medium containing the cells and is moved through the connecting channel 23A, the third culture tank 21AC, and the discharge channel 26A to the discharge container 50.

[0163] The culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the second culture tank 21BB. The cells in the second culture tank 21BB are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23B into the second intermediate tank 60BB. The fluid such as gas in the second intermediate tank 60BB is pushed by the culture medium containing the cells and is moved through the connecting channel 23B, the third culture tank 21BC, and the discharge channel 26B to the discharge container 50.

[0164] The culture medium, moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the second culture tank 21CB. The cells in the second culture tank 21CB are dispersed in the culture medium. The culture medium containing the cells is moved through the connecting channel 23C into the second intermediate tank 60CB. The fluid, such as gas, in the second intermediate tank 60CB is pushed by the culture medium containing the cells and is moved through the connecting channel 23C, the third culture tank 21CC, and the discharge channel 26C to the discharge container 50.

[0165] When culture medium containing cells is placed in the second intermediate tanks 60AB, 60BB, and 60CB of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, which stops moving the fluid.

[0166] For example, after the cell density in the culture medium becomes uniform in each of the second intermediate tanks 60AB, 60BB, and 60CB, the control unit 301 controls the fluid machine 31, which moves the culture medium in the first fluid container 11A towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0167] The culture medium, moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A, reaches the second intermediate tank 60AB. The culture medium containing cells in the second intermediate tank 60AB is moved through the connecting channel 23A into the third culture tank 21AC. The fluid such as gas in the third culture tank 21AC is pushed by the culture medium containing cells and moved through the discharge channel 26A to the discharge container 50.

[0168] The culture medium, moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B, reaches the second intermediate tank 60BB. The culture medium containing cells in the second intermediate tank 60BB is moved through the connecting channel 23B into the third culture tank 21BC. The fluid such as gas in the third culture tank 21BC is pushed by the culture medium containing cells and moved through the discharge channel 26B to the discharge container 50.

[0169] The culture medium, moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C, reaches the second intermediate tank 60CB. The culture medium containing cells in the second intermediate tank 60CB is moved into the third culture tank 21CC via the connecting channel 23C. The fluid, such as gas, in the third culture tank 21CC is pushed by the culture medium containing cells and moved to the discharge container 50 via the discharge channel 26C.

[0170] (Third embodiment) The culture system according to the third embodiment may further include a fifth fluid container 11E, as shown in Figure 4. For example, the fifth fluid container 11E contains factors. For example, the volume of the fifth fluid container 11E is variable. For example, the fifth fluid container 11E is connected to a confluence channel 32. The fifth fluid container 11E may be aseptically connected to the confluence channel 32. The confluence channel 32 is provided with a confluence valve 34E corresponding to the fifth fluid container 11E.

[0171] When delivering factors from the fifth fluid container 11E to the distribution channel 30 via the confluence channel 32, the confluence valve 34E corresponding to the fifth fluid container 11E is opened, and the other confluence valves 33, 34A, 34B, 34C, and 34D are closed. Other components of the culture system according to the third embodiment are the same as those in the first embodiment.

[0172] A factor is, for example, a factor for inducing a cell in a second state, which is different from the first state, from a cell in a first state. The factor may be a nucleic acid such as DNA, RNA, and oligonucleotides, or it may be a protein, a compound, or a virus. DNA may be plasmid DNA. RNA may be mRNA, siRNA, and miRNA. RNA may be modified RNA or unmodified RNA. Viruses may be Sendai virus and lentivirus.

[0173] In this disclosure, induction refers to reprogramming, reprogramming, transformation, transdifferentiation (or lineage reprogramming), differentiation induction, and cell fate reprogramming. Factors that induce cells other than pluripotent stem cells into pluripotent stem cells are called reprogramming factors. Reprogramming factors include, for example, OCT3 / 4, SOX2, KLF4, and c-MYC. Factors that induce stem cells into differentiated cells are called differentiation induction factors.

[0174] Examples of cells in the first state include, but are not limited to, mononuclear cells, stem cells, fibroblasts, nerve cells, retinal epithelial cells, hepatocytes, β-cells, kidney cells, mesenchymal stem cells, blood cells, megakaryocytes, T cells, chondrocytes, cardiomyocytes, muscle cells, vascular cells, epithelial cells, pluripotent stem cells, ES cells, and iPS cells.

[0175] Examples of cells in the second state include, but are not limited to, mononuclear cells, stem cells, fibroblasts, nerve cells, retinal epithelial cells, hepatocytes, β-cells, kidney cells, mesenchymal stem cells, blood cells, megakaryocytes, T cells, chondrocytes, cardiomyocytes, muscle cells, vascular cells, epithelial cells, pluripotent stem cells, ES cells, and iPS cells.

[0176] The factors in the fifth fluid container 11E are distributed to multiple culture devices 20A, 20B, and 20C by the fluid machine 31 via the confluence channel 32 and the distribution channel 30. For example, the type and amount of factors distributed to the multiple culture devices 20A, 20B, and 20C are the same.

[0177] Next, a method for introducing factors into cells using the culture system according to the third embodiment will be described. This method is carried out in appropriate combination with the cell culture method according to the first embodiment.

[0178] When cells are being cultured in the first culture tanks 21AA, 21BA, and 21CA, and factors are to be delivered from the fifth fluid container 11E to the cells in the first culture tanks 21AA, 21BA, and 21CA, the control unit 301 opens the first supply valve 24AA and the first discharge valve 27AA of the first culture device 20A, and closes the other supply valves 24AB, 24AC and the other discharge valves 27AB, 27AC. The control unit 301 opens the first supply valve 24BA and the first discharge valve 27BA of the second culture device 20B, and closes the other supply valves 24BB, 24BC and the other discharge valves 27BB, 27BC. The control unit 301 opens the first supply valve 24CA and the first discharge valve 27CA of the third culture device 20C, and closes the other supply valves 24CB, 24CC and the other discharge valves 27CB, 27CC. Furthermore, the control unit 301 opens the confluence valve 34E and closes the other confluence valves 33, 34A, 34B, 34C, and 34D, and controls the fluid machine 31, which moves the factors in the fifth fluid container 11E towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0179] Factors moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A reach the first culture tank 21AA. Excess fluid in the first culture tank 21AA is moved to the discharge container 50 via the discharge channel 26A.

[0180] Factors moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B reach the first culture tank 21BA. Excess fluid in the first culture tank 21BA is moved to the discharge container 50 via the discharge channel 26B.

[0181] Factors moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C reach the first culture tank 21CA. Excess fluid in the first culture tank 21CA is moved to the discharge container 50 via the discharge channel 26C.

[0182] The length of the distribution channel 30 and / or supply channel 22 may be set so that the factor reaches the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the fifth fluid container 11E to each of the first culture tanks 21AA, 21BA, and 21CA are the same. Alternatively, the timing of the opening of the first supply valves 24AA, 24BA, and 24CA may be made different so that the factor reaches the first culture tanks 24AA, 24BA, and 24CA of the first to third culture devices 20A, 20B, and 20C at different times.

[0183] When the factor is introduced into the first culture tanks 21AA, 21BA, and 21CA of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, which stops moving the fluid. In each of the first culture tanks 21AA, 21BA, and 21CA, the cells are brought into contact with the factor for a predetermined time to introduce the factor into the cells. After that, the culture medium in the first culture tanks 21AA, 21BA, and 21CA may be replaced as described in the first embodiment.

[0184] When cells are being cultured in the second culture tanks 21AB, 21BB, and 21CB, and factors are to be delivered from the fifth fluid container 11E to the cells in the second culture tanks 21AB, 21BB, and 21CB, the control unit 301 opens the second supply valve 24AB and the second discharge valve 27AB of the first culture device 20A, and closes the other supply valves 24AA, 24AC and the other discharge valves 27AA, 27AC. The control unit 301 opens the second supply valve 24BB and the second discharge valve 27BB of the second culture device 20B, and closes the other supply valves 24BA, 24BC and the other discharge valves 27BA, 27BC. The control unit 301 opens the second supply valve 24CB and the second discharge valve 27CB of the third culture device 20C, and closes the other supply valves 24CA, 24CC and the other discharge valves 27CA, 27CC. Furthermore, the control unit 301 opens the confluence valve 34E and closes the other confluence valves 33, 34A, 34B, 34C, and 34D, and controls the fluid machine 31, which moves the factors in the fifth fluid container 11E towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0185] Factors moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A reach the second culture tank 21AB. Excess fluid in the second culture tank 21AB is moved to the discharge container 50 via the discharge channel 26A.

[0186] Factors moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B reach the second culture tank 21BB. Excess fluid in the second culture tank 21BB is moved to the discharge container 50 via the discharge channel 26B.

[0187] Factors moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C reach the second culture tank 21CB. Excess fluid in the second culture tank 21CB is moved to the discharge container 50 via the discharge channel 26C.

[0188] The length of the distribution channel 30 and / or supply channel 22 may be set so that the factor reaches the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the fifth fluid container 11E to each of the second culture tanks 21AB, 21BB, and 21CB are the same. 2 The timing at which the supply valves 24AB, 24BB, and 24CB open is different, and the factor is the first to third culture devices 20A, 20B, and 20C 2 The cells may be allowed to reach culture tanks 24AB, 24BB, and 24CB at different times.

[0189] When the factor is introduced into the second culture tanks 21AB, 21BB, and 21CB of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, causing the fluid machine 31 to stop moving the fluid. In each of the second culture tanks 21AB, 21BB, and 21CB, the cells are brought into contact with the factor for a predetermined time to introduce the factor into the cells. After that, the culture medium in the second culture tanks 21AB, 21BB, and 21CB may be replaced as described in the first embodiment.

[0190] When cells are being cultured in the third culture tanks 21AC, 21BC, and 21CC, and factors are to be delivered from the fifth fluid container 11E to the cells in the third culture tanks 21AC, 21BC, and 21CC, the control unit 301 opens the third supply valve 24AC and the third discharge valve 27AC of the first culture device 20A, and closes the other supply valves 24AA, 24AB and the other discharge valves 27AA, 27AB. The control unit 301 opens the third supply valve 24BC and the third discharge valve 27BC of the second culture device 20B, and closes the other supply valves 24BA, 24BB and the other discharge valves 27BA, 27BB. The control unit 301 opens the third supply valve 24CC and the third discharge valve 27CC of the third culture device 20C, and closes the other supply valves 24CA, 24CB and the other discharge valves 27CA, 27CB. Furthermore, the control unit 301 opens the confluence valve 34E and closes the other confluence valves 33, 34A, 34B, 34C, and 34D, and controls the fluid machine 31, which moves the factors in the fifth fluid container 11E towards the first to third culture devices 20A, 20B, and 20C via the confluence channel 32 and the distribution channel 30.

[0191] Factors moved from the distribution channel 30 to the supply channel 22A of the first culture apparatus 20A reach the third culture tank 21AC. Excess fluid in the third culture tank 21AC is moved to the discharge container 50 via the discharge channel 26A.

[0192] Factors moved from the distribution channel 30 to the supply channel 22B of the second culture apparatus 20B reach the third culture tank 21BC. Excess fluid in the third culture tank 21BC is moved to the discharge container 50 via the discharge channel 26B.

[0193] Factors moved from the distribution channel 30 to the supply channel 22C of the third culture apparatus 20C reach the third culture tank 21CC. Excess fluid in the third culture tank 21CC is moved to the discharge container 50 via the discharge channel 26C.

[0194] The length of the distribution channel 30 and / or supply channel 22 may be set so that the factor reaches the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C simultaneously. For example, the length of the distribution channel 30 and / or supply channel 22 may be set so that the channel lengths from the fifth fluid container 11E to each of the third culture tanks 21AC, 21BC, and 21CC are the same. 3 The timing at which the supply valves 24AC, 24BC, and 24CC open is different, and the factors are controlled in the first to third culture devices 20A, 20B, and 20C. 3 The cultures may be arranged to reach culture tanks 24AC, 24BC, and 24CC at different times.

[0195] When the factor is introduced into the third culture tanks 21AC, 21BC, and 21CC of the first to third culture devices 20A, 20B, and 20C, the control unit 301 controls the fluid machine 31, causing the fluid machine 31 to stop moving the fluid. In each of the third culture tanks 21AC, 21BC, and 21CC, the cells are brought into contact with the factor for a predetermined time to introduce the factor into the cells. After that, the culture medium in the third culture tanks 21AC, 21BC, and 21CC may be replaced as described in the first embodiment.

[0196] The introduction of factors into cells may be carried out in any one or more of the first to third culture tanks 21.

[0197] (Fourth Embodiment) The culture system according to the fourth embodiment may include a plurality of cell containers 10A, 10B, and 10C, as shown in Figure 5. The number of cell containers is not limited. The cells contained in each of the plurality of cell containers 10A, 10B, and 10C are not particularly limited. The cells contained in the plurality of cell containers 10A, 10B, and 10C may be different from each other. The confluence channel 32 is provided with a plurality of confluence valves 33A, 33B, and 33C, each corresponding to the plurality of cell containers 10A, 10B, and 10C.

[0198] When transferring cells from the cell container 10A to the distribution channel 30 via the confluence channel 32, the confluence valve 33A corresponding to the cell container 10A is opened, and the other confluence valves 33B, 33C, 34A, 34B, 34C, and 34D are closed. The cells in the cell container 10A may be distributed to each of the first to third culture devices 20A to 20C, or they may be sent to at least one of the first to third culture devices 20A to 20C.

[0199] When transferring cells from the cell container 10B to the distribution channel 30 via the confluence channel 32, the confluence valve 33B corresponding to the cell container 10B is opened, and the other confluence valves 33A, 33C, 34A, 34B, 34C, and 34D are closed. The cells in the cell container 10B may be distributed to each of the first to third culture devices 20A to 20C, or they may be sent to at least one of the first to third culture devices 20A to 20C.

[0200] When transferring cells from the cell container 10C to the distribution channel 30 via the confluence channel 32, the confluence valve 33C corresponding to the cell container 10C is opened, and the other confluence valves 33A, 33B, 34A, 34B, 34C, and 34D are closed. The cells in the cell container 10C may be distributed to each of the first to third culture devices 20A to 20C, or they may be sent to at least one of the first to third culture devices 20A to 20C.

[0201] Other components of the culture system according to the fourth embodiment are the same as those of at least one of the culture systems according to the first to third embodiments.

[0202] (First example of use of the culture system according to the embodiment) Cells are placed in the cell container 10. Examples of cells include, but are not limited to, mononuclear cells, stem cells, fibroblasts, nerve cells, retinal epithelial cells, hepatocytes, β cells, kidney cells, mesenchymal stem cells, blood cells, megakaryocytes, T cells, chondrocytes, cardiomyocytes, muscle cells, vascular cells, epithelial cells, pluripotent stem cells, ES cells, and iPS cells.

[0203] The concentration of cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, in each of the multiple culture devices 20, cells are grown from single cells in the first culture tank 21. Furthermore, cells are grown in the second and third culture tanks 21. This establishes cell clones in each of the multiple culture devices 20.

[0204] (Second example of use of the culture system according to the embodiment) Factors for inducing cells from the first state to the second state are introduced into the cells in the first state, and these cells are placed in a cell container 10. The concentration of the factor-introduced cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, in each of the multiple culture devices 20, cells from the first state to the second state are induced in the first culture tank 21, and the cells in the second state are cultured on an expansion basis in the second and third culture tanks 21. This establishes clones of the induced cells in each of the multiple culture devices 20.

[0205] (Third example of use of the culture system according to the embodiment) Cells are genetically edited and placed in a cell container 10. For example, cells may be genetically edited by introducing a protein into them. The protein may be a nuclease protein such as Cas9 protein. The concentration of genetically edited cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, in each of the multiple culture devices 20, cells are grown from single cells in the first culture tank 21. Furthermore, cells are grown in the second and third culture tanks 21. In this way, clones of genetically edited cells are established in each of the multiple culture devices 20.

[0206] (Fourth example of use of the culture system according to the embodiment) Stem cells are placed in a cell container 10. Factors for inducing somatic cells from stem cells are placed in a fifth fluid container 11E. The concentration of stem cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, in each of the multiple culture devices 20, stem cells are cultured in suspension in the first culture tank 21 to form spheroids, factors are introduced into the spheroids in the second culture tank 21, and the cells with the introduced factors are cultured in adhesion in the third culture tank 21 to induce somatic cells. In this way, somatic cells derived from single cells are established in each of the multiple culture devices 20.

[0207] (Fifth example of use of the culture system according to the embodiment) Stem cells are placed in the cell container 10. Factors for inducing somatic cells from stem cells are placed in the fifth fluid container 11E. The concentration of stem cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, in each of the multiple culture devices 20, stem cells are cultured in adherent culture or suspension culture in the first culture tank 21, and factors are introduced into the stem cells. In the second and third culture tanks 21, the induced cells are cultured in adherent culture or suspension culture, and the induced cells are proliferated. In this way, induced somatic cells derived from single cells are established in each of the multiple culture devices 20.

[0208] (Sixth example of use of the culture system according to the embodiment) Somatic cells are placed in the cell container 10. Factors for inducing stem cells from somatic cells are placed in the fifth fluid container 11E. The concentration of somatic cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, in each of the multiple culture devices 20, somatic cells are cultured in adherent culture or suspension culture in the first culture tank 21, and factors are introduced into the somatic cells. In the second and third culture tanks 21, the induced cells are cultured in adherent culture or suspension culture, and the induced cells are grown. In this way, single-cell derived induced stem cells are established in each of the multiple culture devices 20.

[0209] (Seventh example of use of the culture system according to the embodiment) Differentiated cells are placed in a cell container 10. Factors for inducing other differentiated cells from the differentiated cells are placed in a fifth fluid container 11E. The concentration of differentiated cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, in each of the multiple culture devices 20, somatic cells are cultured in adherent or suspension culture in the first culture tank 21, and factors are introduced into the differentiated cells. In the second and third culture tanks 21, the differentiated cells are cultured in adherent or suspension culture, and the differentiated cells are grown. This allows for lineage-specific differentiation induction in each of the multiple culture devices 20.

[0210] (An eighth example of use of the culture system according to the embodiment) Different chimeric antigen receptor (CAR) gene-modified T cells (CAR-T cells) are placed in a cell container 10. The concentration of CAR-T cells placed in the cell container 10 is set so that single cells are distributed to each of the multiple culture devices 20. For example, single cells of the first type of CAR-T cell are distributed to the first culture device 20A, single cells of the second type of CAR-T cell are distributed to the second culture device 20B, and single cells of the third type of CAR-T cell are distributed to the third culture device 20A. For example, in each of the multiple culture devices 20, the cells are grown from single cells in the first culture tank 21. Furthermore, the cells are grown in the second and third culture tanks 21. This establishes clones of CAR-T cells in each of the multiple culture devices 20.

[0211] As described above, the present invention has been described by embodiments, but the descriptions and drawings that form part of this disclosure should not be understood as limiting the invention. Various alternative embodiments, configurations, and operational techniques should become apparent to those skilled in the art from this disclosure. For example, if the supply valve 24AB and the discharge valve 27AB are closed, even if the connecting valve 25AA is open, the fluid in the culture tank 21AA may not be able to enter the connecting passage 23A due to atmospheric pressure. Therefore, even if the valve is not closed, if the fluid cannot pass through the valve due to pressure, etc., it is not necessary to close the valve. Thus, it should be understood that the present invention encompasses various embodiments and the like that are not described herein. [Explanation of symbols]

[0212] 10...Cell container, 11A, 11B, 11C, 11D, 11E...Fluid container, 20, 20A, 20B, 20C...Culture device, 21, 21AA, 21AB, 21AC, 21BA, 21BB, 21BC, 21CA, 21CB, 21CC...Culture tank, 22, 22A, 22B, 22 C... Supply channel, 23, 23A, 23B, 23C... Connection channel, 24, 24AA, 24AB, 24AC, 24BA, 24BB, 24BC, 24CA, 24CB, 24CC... Supply valve, 25, 25AA, 25AB, 25BA, 25BB, 25CA, 25CB... Connection valve, 26, 26 A, 26B, 26C... Discharge channels, 27, 27AA, 27AB, 27AC, 27BA, 27BB, 27BC, 27CA, 27CB, 27CC... Discharge valves, 28, 28A, 28B, 28C... Recovery valves, 30... Distribution channels, 31... Fluid machinery, 32... Confluence channels, 33, 34A, 34B, 34C, 34D, 34E... Confluence valves, 40, 40A, 40B, 40C... Cell recovery channels, 41, 41A, 41B, 41C... Cell recovery containers, 50... Discharge containers, 60, 60AA, 60AB, 60BA, 60BB, 60CA, 60CB... Intermediate tanks, 301... Control unit

Claims

1. A cell containment container configured to house cells, Multiple culture devices, A distribution channel configured to distribute the cells from the cell container to the plurality of culture devices, A culture system comprising, Each of the aforementioned multiple culture devices, Multiple culture tanks, A supply channel configured to selectively supply fluid from the distribution channel to the plurality of culture tanks, A connecting channel configured to connect the plurality of culture tanks, Equipped with, In each of the plurality of culture devices, the plurality of culture tanks are connected in series by the connecting channel, In each of the plurality of culture devices, the supply channel is configured to selectively supply fluid to the plurality of culture tanks via the connecting channel, so that cells move sequentially between the plurality of culture tanks. Culture system.

2. The culture system according to claim 1, further comprising a fluid machine configured to supply fluid from the distribution channel to the supply channel of each of the plurality of culture devices.

3. The culture system according to claim 1 or 2, wherein each of the plurality of culture devices further comprises a plurality of supply valves provided in the supply channel.

4. The culture system according to any one of claims 1 to 3, wherein each of the plurality of culture devices further comprises a connecting valve provided in the connecting channel.

5. The culture system according to any one of claims 1 to 4, wherein each of the plurality of culture devices further comprises an intermediate tank provided in the connecting channel.

6. The culture system according to any one of claims 1 to 5, wherein each of the plurality of culture devices further comprises a discharge channel configured to selectively discharge fluid from the plurality of culture tanks.

7. The culture system according to claim 6, wherein in each of the plurality of culture devices, the discharge channel is configured to selectively discharge fluid from the plurality of culture tanks so that the cells move sequentially through the connecting channel.

8. Each of the aforementioned multiple culture devices, Multiple supply valves provided in the supply channel, A discharge channel configured to selectively discharge fluid from the plurality of culture tanks, Multiple discharge valves provided in the aforementioned discharge channel, Furthermore, In each of the plurality of culture devices, the plurality of supply valves each correspond to the plurality of culture tanks, In each of the plurality of culture devices, the plurality of discharge valves each correspond to the plurality of culture tanks, One of the supply valves corresponding to one of the culture tanks from which the cells are supplied from the cell containment container opens. One of the multiple discharge valves corresponding to one of the multiple culture tanks from which the cells are supplied from the cell containment container opens. The culture system according to claim 1 or 2.

9. Each of the aforementioned multiple culture devices, Multiple supply valves provided in the supply channel, A discharge channel configured to selectively discharge fluid from the plurality of culture tanks, Multiple discharge valves provided in the aforementioned discharge channel, Furthermore, In each of the plurality of culture devices, the plurality of supply valves each correspond to the plurality of culture tanks, In each of the plurality of culture devices, the plurality of discharge valves each correspond to the plurality of culture tanks, One of the supply valves corresponding to one of the culture tanks to which the fluid is supplied opens, One of the multiple discharge valves corresponding to one of the multiple culture tanks to which the fluid is supplied opens. The culture system according to claim 1 or 2.

10. Each of the aforementioned multiple culture devices, Multiple supply valves provided in the supply channel, Multiple connecting valves provided in the aforementioned connecting passage, A discharge channel configured to selectively discharge fluid from the plurality of culture tanks, Multiple discharge valves provided in the aforementioned discharge channel, Furthermore, In each of the plurality of culture devices, the plurality of supply valves each correspond to the plurality of culture tanks, In each of the above-mentioned culture apparatuses, the above-mentioned connecting valves are provided between the above-mentioned culture tanks. In each of the plurality of culture devices, the plurality of discharge valves each correspond to the plurality of culture tanks, One of the supply valves corresponding to one of the culture tanks in which the cells are arranged opens, The connecting valve between the culture tank in which the cells are placed and the culture tank adjacent to the culture tank in which the cells are placed opens. One of the multiple discharge valves corresponding to the adjacent culture tank opens. The culture system according to claim 1 or 2.

11. The system further comprises one or more fluid containers configured to contain a fluid, The distribution channel is configured to distribute the fluid from the fluid container to the plurality of culture devices. A culture system according to any one of claims 1 to 10.

12. The culture system according to claim 11, wherein the fluid comprises a culture medium, a buffer, a cell delaminating agent, a cryopreservation agent, or a factor.

13. A method for culturing cells, A culture system comprising a cell container configured to contain cells, a plurality of culture devices, and a distribution channel configured to distribute the cells from the cell container to the plurality of culture devices, wherein each of the plurality of culture devices comprises a plurality of culture tanks, a supply channel configured to selectively supply fluid from the distribution channel to the plurality of culture tanks, and a connecting channel configured to connect the plurality of culture tanks, wherein in each of the plurality of culture devices, the plurality of culture tanks are connected in series by the connecting channel, and in each of the plurality of culture devices, the supply channel is configured to selectively supply fluid to the plurality of culture tanks via the connecting channel so that the cells move sequentially through the plurality of culture tanks. Distributing the cells from the cell container to each of the plurality of culture devices via the distribution channel, In each of the plurality of culture devices, the cells are supplied to at least one of the plurality of culture tanks via the supply channel. In each of the plurality of culture devices, the cells are cultured in at least one of the plurality of culture tanks. A method for culturing cells, including the following.

14. Each of the plurality of culture devices further comprises a discharge channel configured to selectively discharge fluid from the plurality of culture tanks, In each of the plurality of culture devices, when supplying cells to at least one of the plurality of culture tanks via the supply channel, the fluid in at least one of the plurality of culture tanks is discharged into the discharge channel. The cell culture method according to claim 13.

15. The cell culture method according to claim 13, further comprising supplying fluid to the culture tank in which the cells are placed, among the plurality of culture vessels, via the supply channel, in each of the plurality of culture devices.

16. Each of the plurality of culture devices further comprises a discharge channel configured to selectively discharge fluid from the plurality of culture tanks, In each of the plurality of culture devices, the fluid is supplied to the culture tank in which the cells are placed, via the supply channel, such that the fluid in the culture tank in which the cells are placed is discharged to the discharge channel. The cell culture method according to claim 15.

17. The cell culture method according to claim 13, further comprising moving the cells from one culture tank in which the cells are located to another culture tank via the connecting channel in each of the plurality of culture devices.

18. Each of the plurality of culture devices further comprises a discharge channel configured to selectively discharge fluid from the plurality of culture tanks, The cell culture method according to claim 17, wherein in each of the plurality of culture devices, when moving the cells from a culture tank in which the cells are placed to another culture tank via the connecting channel, the fluid in the other culture tank is discharged into the discharge channel.

19. The cell culture method according to any one of claims 13 to 18, wherein the fluid is a gas, a liquid, or a gel.

20. The cell culture method according to any one of claims 13 to 19, wherein the fluid contains a factor, and in each of the plurality of culture devices, in at least one of the plurality of culture tanks, the factor induces cells in a different state from the cells.

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