Cell culture system and cell culture method

Abstract

To provide a cell culture system that, even in the case where gas is mixed into a flow channel connecting the liquid feeding part and the cell culture device when connecting a liquid feeding part and a cell culture device, is capable of preventing gas in a flow channel connecting the liquid feeding part and the cell culture device from flowing into the cell culture device.SOLUTION: The cell culture system 100 comprises: a cell culture device 1 having a culture chamber 1a capable of housing a cell 30 and cell culture solution 31; a liquid feeding part 2 which feeds the cell culture solution 31 to the culture chamber 1a; a first culture liquid flow channel 3 connecting the culture chamber 1a and the liquid feeding part 2; a branch flow channel 4 which is provided while branched from the first culture liquid flow channel 3, and used for removing gas 32 in the first culture liquid flow channel 3; a flow channel switching part 5 which switches a first state 50 that the cell culture solution 31 is discharged via the branch flow channel 4 and a second state 51 that the cell culture solution 31 flows into the culture chamber 1a.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a cell culture system and a cell culture method. [Background technology]

[0002] BACKGROUND ART Conventionally, a cell culture system and a cell culture method are known (see, for example, Patent Document 1).

[0003] The cell culture system disclosed in Patent Document 1 includes a cell culture substrate, a liquid supply pump, and a waste liquid tank. The liquid supply pump and the cell culture substrate are connected via a liquid supply pad, and the cell culture system discloses a cell culture medium that is pumped from the liquid supply pump and supplied to the cell culture substrate. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2013 / 011962 Summary of the Invention [Problem to be solved by the invention]

[0005] Here, when a culture medium is sent from a liquid delivery unit (liquid delivery pump) to a cell culture device (cell culture substrate) as in the cell culture system disclosed in Patent Document 1, if gas is present in the piping connecting the liquid delivery unit and the cell culture device, the gas will enter the cell culture device when the culture medium is sent from the liquid delivery unit to the cell culture device. If gas enters the cell culture device, the seeded cells will come into contact with the gas. If the cells come into contact with the gas, there is the inconvenience that the cells may be damaged.

[0006] In order to avoid gas getting into the cell culture device, it is necessary to fill the piping connecting the liquid delivery unit and the cell culture device with culture medium before sending the culture medium from the liquid delivery unit to the cell culture device.

[0007] One possible method for achieving this is to pump the culture medium from the liquid delivery unit to the end of the piping before connecting the piping to the cell culture device, and then push out any gas that may have been present in the piping from the end of the piping. However, this method has the problem that it is difficult to connect the piping to the cell culture device without introducing gas into the piping or the cell culture device when the piping connecting the liquid delivery unit to the cell culture device is filled with culture medium up to the end. Furthermore, the connection of the piping to the cell culture device must be performed in a sterile space to prevent contamination, which reduces the convenience of the cell culture system.

[0008] A second method is to install a mechanism for removing gas that has been present in the piping connecting the piping to the cell culture device. In this case, the piping can be filled with culture medium. However, there is a problem in that gas still remains in the piping connecting the gas removal mechanism to the cell culture device.

[0009] This invention has been made to solve the above-mentioned problems, and one object of this invention is to provide a cell culture system and a cell culture method that can prevent gas in the flow path connecting the liquid delivery unit and the cell culture device from entering the cell culture device, even if gas gets mixed into the flow path connecting the liquid delivery unit and the cell culture device when connecting the liquid delivery unit and the cell culture device. [Means for solving the problem]

[0010] In order to achieve the above object, a cell culture system according to a first aspect of the present invention includes a cell culture device having a culture chamber capable of accommodating cells and a cell culture solution; Located outside the cell culture device, a fluid delivery unit that delivers a cell culture solution to the culture chamber; a portion of which is disposed within a cell culture device;a first culture solution flow path connecting the culture chamber and the fluid supply unit; a branch flow path branching from the first culture solution flow path for removing gas in the first culture solution flow path; a flow path switching unit switching between a first state in which the cell culture solution is discharged through the branch flow path and a second state in which the cell culture solution enters the culture chamber; a cell seeding flow path connected to the first culture solution flow path and through which a cell suspension flows; and a control unit controlling the switching between the first state and the second state by the flow path switching unit, wherein the branching portion of the first culture solution flow path and the branch flow path is located in the first culture solution flow path in the cell culture device closer to the culture chamber than the connection portion of the cell seeding flow path, and the control unit In the first state, after the cell culture medium starts to be sent into the first culture medium flow path, a predetermined time has elapsed, With the cell suspension filling the portion on the culture chamber side of the branching portion, the flow path switching unit switches from the first state to the second state.

[0011] In order to achieve the above object, a cell culture method according to a second aspect of the present invention is a cell culture method for culturing cells using a cell culture device having a culture chamber capable of accommodating cells and a cell culture solution, the method comprising the steps of seeding cells in the culture chamber; It is located outside the cell culture device. A fluid delivery unit that delivers cell culture fluid to the culture chamber is connected. and a part of it is installed in the cell culture device. The method includes the steps of: removing gas in the first culture solution flow path by pumping the cell culture solution in a first state in which the cell culture solution is discharged through a branch flow path branched from the first culture solution flow path; switching from the first state to a second state in which the cell culture solution enters a culture chamber; and pumping the cell culture solution in the second state, wherein the branching portion of the first culture solution flow path and the branch flow path is located in the first culture solution flow path in the cell culture device closer to the culture chamber than the connecting portion of the cell seeding flow path connected to the first culture solution flow path and through which the cell suspension flows; and in the step of switching from the first state to the second state, In the first state, after the cell culture medium starts to be sent into the first culture medium flow path, a predetermined time has elapsed, When the cell suspension fills the part on the culture chamber side of the branch part, the first state is switched to the second state. [Effects of the Invention]

[0012] In the cell culture system according to the first aspect of the present invention, as described above, even if gas is mixed into the first culture solution flow path when connecting the fluid delivery unit and the first culture solution flow path, the gas in the first culture solution flow path can be removed through the branch flow path by discharging the cell culture solution from the branch flow path in the first state. As a result, unlike a configuration in which the first culture solution flow path and the cell culture device are connected in a state in which the first culture solution flow path is filled with cell culture solution up to its end, a cell culture system can be provided that can prevent gas in the flow path connecting the fluid delivery unit and the cell culture device from entering the cell culture device when connecting the fluid delivery unit and the cell culture device.

[0013] Furthermore, a cell culture method according to a second aspect of the present invention includes the steps of removing gas from the first culture solution flow path, switching from the first state to a second state in which the cell culture solution enters the culture chamber, and delivering the cell culture solution in the second state, as described above. This provides a cell culture method that can prevent gas in the flow path connecting the solution delivery unit and the cell culture device from entering the cell culture device, even if gas gets mixed into the flow path connecting the solution delivery unit and the cell culture device when connecting the solution delivery unit and the cell culture device, just like the cell culture system according to the first aspect. [Brief explanation of the drawings]

[0014] [Figure 1] 1 is a block diagram showing the configuration of a cell culture system according to one embodiment. [Figure 2] FIG. 1 is a schematic diagram of a cell culture device according to one embodiment. [Figure 3] FIG. 3 is a cross-sectional view taken along line 200-200 in FIG. 2. [Figure 4] FIG. 2 is a schematic diagram illustrating a first state according to one embodiment. [Figure 5] FIG. 4 is a schematic diagram illustrating a second state according to one embodiment. [Figure 6]FIG. 2 is a schematic diagram illustrating the connection between the cell culture device and each flow channel. [Figure 7] FIG. 1 is a schematic diagram illustrating a process of seeding cells into a cell culture device. [Figure 8] FIG. 10 is a schematic diagram illustrating a configuration in which a liquid delivery unit and a cell culture device are connected. [Figure 9] FIG. 10 is a schematic diagram illustrating the movement of gas present in a first cell culture solution when the cell culture solution is delivered from a solution delivery unit. [Figure 10] FIG. 10 is a schematic diagram illustrating a configuration in which gas in a first culture solution flow path is caused to flow into a branch flow path. [Figure 11] FIG. 11 is an enlarged view of a partial area of ​​FIG. [Figure 12] FIG. 10 is a schematic diagram illustrating a configuration in which the cell culture fluid delivered from the fluid delivery section flows into a first discharge flow path. [Figure 13] FIG. 10 is a schematic diagram illustrating a configuration in which the cell culture medium is discharged into a culture medium reservoir via a first discharge flow path and a flow path switching unit. [Figure 14] FIG. 2 is a schematic diagram of a cell culture system in a second state according to one embodiment. [Figure 15] FIG. 10 is a schematic diagram illustrating a configuration in which the cell culture medium is discharged into a culture medium reservoir via a second discharge flow path and a flow path switching unit. [Figure 16] 1 is a flowchart illustrating a cell culture process according to one embodiment. [Figure 17] FIG. 2 is a schematic diagram illustrating the configuration of a cell culture system according to a first modified example and a first state. [Figure 18] FIG. 10 is a schematic diagram for explaining a second state of the cell culture system according to the first modified example. [Figure 19] FIG. 10 is a schematic diagram for explaining the configuration of a cell culture system according to a second modified example. [Figure 20] FIG. 10 is a cross-sectional view illustrating the configuration of a cell culture device according to a third modified example. [Figure 21] FIG. 10 is a block diagram showing the configuration of a cell culture system according to a fourth modified example. DETAILED DESCRIPTION OF THE INVENTION

[0015] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, specific embodiments of the present invention will be described with reference to the accompanying drawings.

[0016] (Configuration of cell culture system) The configuration of a cell culture system 100 according to one embodiment will be described with reference to FIGS.

[0017] As shown in Figure 1, the cell culture system 100 includes a cell culture device 1, a fluid delivery section 2, a first culture solution flow path 3, a branch flow path 4, a flow path switching section 5, a control section 6, a culture solution storage section 7, and a memory section 8.

[0018] The cell culture device 1 has a culture chamber 1a. The cell culture device 1 is a device for culturing seeded cells 30. The cell culture device 1 includes a cell culture vessel made of, for example, resin or glass. The detailed configuration of the cell culture device 1 will be described later.

[0019] The culture chamber 1a is configured to be able to accommodate cells 30 and a cell culture solution 31. The culture chamber 1a is a storage space formed to be able to accommodate the cells 30 and the cell culture solution 31.

[0020] The liquid delivery unit 2 is configured to deliver the cell culture solution 31 to the culture chamber 1a. The liquid delivery unit 2 includes, for example, a syringe pump. Details of the configuration of the liquid delivery unit 2 to deliver the cell culture solution 31 will be described later.

[0021] The first culture solution flow path 3 includes a first groove 3a and a first liquid supply tube 3b. The first groove 3a and the first liquid supply tube 3b are connected at a first opening 3c (see FIG. 2). The first groove 3a is a groove formed integrally with the cell culture device 1. The first liquid supply tube 3b is provided outside the cell culture device 1. The first liquid supply tube 3b is configured to connect the liquid supply section 2 and the cell culture device 1. The first liquid supply tube 3b includes, for example, a liquid supply tube made of silicone. The detailed configuration of the first culture solution flow path 3 will be described later.

[0022] The branch flow path 4 branches off from the first culture solution flow path 3. The branch flow path 4 includes a second groove 4a and a second liquid supply tube 4b. The second groove 4a and the second liquid supply tube 4b are connected at a second opening 4c (see FIG. 2). The second groove 4a is a groove formed integrally with the cell culture device 1. In this embodiment, the second groove 4a branches off from the first groove 3a. The second liquid supply tube 4b is provided outside the cell culture device 1. The second groove 4a connects the second groove 4a and the flow path switching unit 5. The second liquid supply tube 4b includes, for example, a liquid supply tube made of silicon. The branch flow path 4 is a flow path for removing gas 32 (see FIG. 8) from the first culture solution flow path 3. Details of the configuration for removing gas 32 from the first culture solution flow path 3 by the branch flow path 4 will be described later.

[0023] The cell culture device 1 also has a second culture solution flow path 9 that connects to the culture chamber 1a at a position different from the first culture solution flow path 3. The second culture solution flow path 9 includes a third groove portion 9a and a third liquid supply tube 9b. The third groove portion 9a and the third liquid supply tube 9b are connected at a third opening portion 9c (see FIG. 2). The third groove portion 9a is a groove portion that is integrally formed with the cell culture device 1. The third liquid supply tube 9b is provided outside the cell culture device 1. The third liquid supply tube 9b connects the third groove portion 9a and the flow path switching unit 5. The third liquid supply tube 9b includes, for example, a liquid supply tube made of silicone.

[0024] The flow path switching unit 5 includes a switching valve 5a and a drive unit 5b. The flow path switching unit 5 is configured to switch between a first state 50 (see FIG. 4) in which the cell culture solution 31 delivered from the fluid delivery unit 2 is discharged via the first culture solution flow path 3 and the branch flow path 4, and a second state 51 (see FIG. 5) in which the cell culture solution 31 delivered from the fluid delivery unit 2 enters the culture chamber 1a. Details of the configuration of the flow path switching unit 5 for switching between the first state 50 and the second state 51 will be described later.

[0025] The switching valve 5a is configured to switch a discharge flow path, which discharges the cell culture solution 31 delivered from the solution delivery section 2 from the cell culture device 1, between a first state 50 and a second state 51. The discharge flow path includes a first discharge flow path 10a formed by the first culture solution flow path 3 and the branch flow path 4, and a second discharge flow path 10b formed by the first culture solution flow path 3, the culture chamber 1a, and the second culture solution flow path 9. The first discharge flow path 10a is formed by the first solution delivery tube 3b, a part of the first groove portion 3a, the second groove portion 4a, and the second solution delivery tube 4b. The second discharge flow path 10b is formed by the first solution delivery tube 3b, the first groove portion 3a, the third groove portion 9a, and the third solution delivery tube 9b.

[0026] The driving unit 5b is configured to drive the switching valve 5a under the control of the control unit 6. The driving unit 5b includes, for example, a stepping motor.

[0027] The control unit 6 is configured to control switching between the first state 50 and the second state 51 by the flow path switching unit 5. The control unit 6 includes, for example, a CPU (Central Processing Unit), a ROM (Read Only Memory), and a RAM (Random Access Memory).

[0028] The culture medium reservoir 7 is connected to the flow path switching unit 5 by the third discharge flow path 10c, and is configured to store the cell culture medium 31 that is discharged to the outside of the cell culture device 1. The culture medium reservoir 7 includes, for example, a glass bottle, a resin tube, or the like.

[0029] The storage unit 8 is configured to store programs and the like executed by the control unit 6. The storage unit 8 includes, for example, an HDD (Hard Disk Drive) or a non-volatile memory.

[0030] (Configuration of cell culture device) 2, the first groove portion 3a is provided in the cell culture device 1. In the cell culture device 1, one end of the first groove portion 3a is connected to the culture chamber 1a, and the other end is connected to the first opening 3c.

[0031] Furthermore, in the cell culture device 1, one end of the third groove portion 9a is connected to the culture chamber 1a, and the other end is connected to the third opening 9c.

[0032] The second groove portion 4a is provided branching off from the first groove portion 3a in the cell culture device 1. One end of the second groove portion 4a is connected to the first groove portion 3a in the cell culture device 1, and the other end is connected to the second opening 4c.

[0033] The cell culture device 1 also has a cell seeding channel 11. The cell seeding channel 11 is a groove portion integrally formed with the cell culture device 1. The cell seeding channel 11 is connected to the first groove portion 3a. The cell seeding channel 11 is a channel for seeding cells 30. One end of the cell seeding channel 11 is connected to the first groove portion 3a, and the other end is connected to the fourth opening 11a.

[0034] The cell culture device 1 also has an exhaust flow path 12. The exhaust flow path 12 is a groove portion integrally formed in the cell culture device 1. The exhaust flow path 12 is connected to the third groove portion 9a. The exhaust flow path 12 is a flow path for discharging gas 32 discharged from the culture chamber 1a when cells 30 are seeded. One end of the exhaust flow path 12 is connected to the third groove portion 9a, and the other end is connected to the fifth opening 12a.

[0035] In this embodiment, the branching portion 3d of the first culture solution flow path 3 is provided at a position in the first culture solution flow path 3 (first groove portion 3a) in the cell culture device 1 closer to the culture chamber 1a than the connection portion 3e of the cell seeding flow path 11. In other words, the second groove portion 4a branches off from a position in the first groove portion 3a between the connection portion 3e and the portion where the culture chamber 1a is connected to the first groove portion 3a. Specifically, the second groove portion 4a branches off from a position where the distance d1 from the end of the culture chamber 1a connected to the first groove portion 3a to the branching portion 3d is smaller than the distance d2 from the end of the culture chamber 1a connected to the first groove portion 3a to the connection portion 3e.

[0036] In this embodiment, cells 30 are cultured in a state of adhering to the bottom surface 1b of the culture chamber 1a, as shown in Fig. 3. The bottom surface 1b of the culture chamber 1a is coated with protein or subjected to plasma treatment, for example, to allow cells 30 to adhere thereto.

[0037] (First and second states) Next, the first state 50 and the second state 51 will be described with reference to FIGS.

[0038] 4, the flow path switching unit 5 is connected to the branch flow path 4 and the second culture solution flow path 9. Specifically, the flow path switching unit 5 is connected to the second liquid supply tube 4b and the third liquid supply tube 9b. The flow path switching unit 5 is configured to switch between a first state 50 and a second state 51 by blocking the end of the branch flow path 4 (the second liquid supply tube 4b) or the end of the second culture solution flow path 9 (the third liquid supply tube 9b).

[0039] As shown in FIG. 4, in this embodiment, the switching valve 5a includes a four-way valve. The switching valve 5a includes four ports, each connected to the second liquid supply tube 4b, the third liquid supply tube 9b, the third discharge flow path 10c, and the fourth discharge flow path 10d, respectively. One end of the fourth discharge flow path 10d is connected to the switching valve 5a, and the other end is blocked by a plug 60. Therefore, the flow path connected to the fourth discharge flow path 10d is blocked by the plug 60. The plug 60 includes, for example, a rubber cap. The control unit 6 is configured to control the drive unit 5b to change the connection state of each port of the switching valve 5a, thereby switching between a first state 50 and a second state 51.

[0040] In the first state 50, the second liquid supply tube 4b is connected to the third discharge flow path 10c and the third liquid supply tube 9b is connected to the fourth discharge flow path 10d by the switching valve 5a. In the example shown in FIG. 4, the connected flow paths are shown with the same hatching to make it easier to understand the connection relationship. Specifically, in the example shown in FIG. 4, the second liquid supply tube 4b and the third discharge flow path 10c are shown with the same hatching. Furthermore, the third liquid supply tube 9b and the fourth discharge flow path 10d are shown with the same hatching.

[0041] As shown in Fig. 5, the second state 51 is a state in which the second liquid supply tube 4b is connected to the fourth discharge flow path 10d and the third liquid supply tube 9b is connected to the third discharge flow path 10c by the switching valve 5a. In the example shown in Fig. 5, similar to the example shown in Fig. 4, the second liquid supply tube 4b and the fourth discharge flow path 10d are illustrated with the same hatching to make the connection relationship easier to understand. Furthermore, the third liquid supply tube 9b and the third discharge flow path 10c are illustrated with the same hatching.

[0042] 6 is a schematic diagram showing the connection state of the first liquid supply tube 3b, the third liquid supply tube 9b, the flow path switching unit 5, and the culture solution storage unit 7 to the cell culture device 1. Before the cells 30 are seeded, the cell culture device 1 is sterilized in advance in the state shown in FIG. 6. Examples of sterilization methods that can be used include sterilization by radiation, sterilization by electron beam, sterilization by high-pressure steam, and dry heat sterilization.

[0043] The grooves of each flow path and the liquid supply tubes have different diameters. The diameter of the grooves of each flow path is smaller than the diameter of the liquid supply tubes of each flow path. The diameter of the grooves of each flow path is, for example, approximately 100 μm. The diameter of the liquid supply tubes of each flow path is, for example, approximately 1 mm. The diameter of the cell seeding flow path 11 and the diameter of the exhaust flow path 12 are, for example, approximately 100 μm.

[0044] (cell seeding) Next, the process of seeding cells 30 in the culture chamber 1a will be described with reference to Fig. 7. In Fig. 7 to Fig. 15, the cell culture solution 31 is shown hatched. That is, in each flow path, the liquid delivery unit 2, and the culture solution storage unit 7, the hatched parts are parts filled with the cell culture solution 31, and the unhatched parts are parts filled with the gas 32.

[0045] In this embodiment, as shown in FIG. 7, cells 30 are seeded in the culture chamber 1a through the cell seeding channel 11. Specifically, a micropipette 20 is used to feed a cell suspension into the cell seeding channel 11 through the fourth opening 11a, thereby seeding the cells 30 in the culture chamber 1a. When seeding the cells 30 through the cell seeding channel 11, as shown in FIG. 7, the end of the first liquid supply tube 3b that is not connected to the first opening 3c is blocked with a plug 61, and each port of the switching valve 5a is disconnected from any channel, so that the cell suspension does not flow outside the culture chamber 1a. After the seeding of the cells 30 is completed, the fourth opening 11a and the fifth opening 12a are blocked with plugs 62 (see FIG. 8) and 63 (see FIG. 8), respectively. Each of the plugs 60, 61, and 62 includes, for example, a rubber cap. The cells 30 are seeded in a sterile space such as a clean bench (not shown). That is, the process shown in FIG. 7 is performed in a sterile space. After the cells 30 are seeded, they are cultured in a thermostatic chamber (not shown), which is a non-sterile space. The thermostatic chamber includes, for example, a CO2 incubator.

[0046] (Replacing the cell culture medium in the culture chamber) Next, a process for replacing the cell culture solution 31 in the culture chamber 1a will be described with reference to Figures 8 to 13. The processes shown in Figures 8 to 13 are carried out in a non-sterile space such as a thermostatic bath.

[0047] In this embodiment, cells 30 are seeded in the culture chamber 1a and then grow by adhering to the bottom surface 1b of the culture chamber 1a. As the cells 30 grow, they acquire nutrients contained in the cell culture solution 31 and excrete waste products generated during the growth. Therefore, in order to culture the cells 30, the cell culture solution 31 needs to be replaced periodically. In this embodiment, after the cells 30 are seeded and the cells 30 adhere to the bottom surface 1b of the culture chamber 1a, the cell culture solution 31 is replaced by being sent by the solution sending unit 2.

[0048] As shown in FIG. 8, the liquid delivery section 2 includes a cylinder 2a, a plunger (pusher) 2b, a drive source 2c, a cylinder holding section 2d, and a plunger holding section 2e.

[0049] The cylinder 2a is configured to store the cell culture solution 31 therein. The cylinder 2a is connected to a first liquid supply tube 3b and is configured to be able to supply the stored cell culture solution 31 to the first liquid supply tube 3b. The plunger 2b is held by a plunger holding unit 2e. A driving source 2c is configured to move the plunger holding unit 2e. The driving source 2c includes, for example, a stepping motor. The plunger holding unit 2e holds the plunger 2b. The plunger holding unit 2e is configured to move the plunger 2b when moved by the driving source 2c. When the plunger holding unit 2e is moved, the plunger 2b moves, and the cell culture solution 31 in the cylinder 2a is discharged from the cylinder 2a. Of the liquid supply unit 2, the cylinder 2a and plunger 2b are sterilized in advance before storing the cell culture solution 31. Furthermore, the cylinder 2a and plunger 2b are discarded every time the delivery of the cell culture solution 31 is completed. That is, the cylinder 2a and plunger 2b are disposable. The fluid delivery unit 2 other than the cylinder 2a and plunger 2b is not disposable and includes the drive source 2c, etc., making it difficult to sterilize them.

[0050] (Removal of gas from the first culture medium flow path) As shown in FIG. 8, after the cells 30 are seeded, when the first liquid supply tube 3b and the liquid supply unit 2 (cylinder 2a) are connected, gas 32 is contained within the first liquid supply tube 3b. If the cell culture solution 31 is sent from the liquid supply unit 2 in this state, the gas 32 will enter the culture chamber 1a. When the gas 32 enters the culture chamber 1a, the cells 30 come into contact with the gas 32. When the cells 30 come into contact with the gas 32, the cells 30 may be damaged or killed. Note that the connection between the first liquid supply tube 3b and the liquid supply unit 2 is performed in a sterile space after the cells 30 are seeded.

[0051] Therefore, in this embodiment, when the first liquid supply tube 3b is filled with cell culture solution 31 up to the end, the gas 32 in the first culture solution flow path 3 is removed by a method different from the method of connecting the first liquid supply tube 3b to the cell culture device 1, and then the cell culture solution 31 is supplied to the culture chamber 1a.

[0052] 9, in a first state 50 in which the cell culture solution 31 is discharged from the first discharge flow path 10a by the flow path switching unit 5, the cell culture solution 31 is delivered from the liquid delivery unit 2. When the delivery of the cell culture solution 31 from the liquid delivery unit 2 starts, the gas 32 in the first liquid delivery tube 3b moves toward the culture chamber 1a.

[0053] In the first state 50, the third liquid supply tube 9b is connected to the fourth discharge flow path 10d. Therefore, as shown in Fig. 10, the cell culture solution 31 supplied from the first liquid supply tube 3b flows into the second groove portion 4a via the first groove portion 3a.

[0054] Fig. 11 is a schematic enlarged view of the circled portion 40 in Fig. 10. As shown in Fig. 11, when cell culture solution 31 is delivered from solution delivery section 2 in first state 50, cell culture solution 31 flows into second groove section 4a.

[0055] Here, in the first groove portion 3a, a portion 3f between the second groove portion 4a and the culture chamber 1a is filled with the cell culture solution 31 when the cells 30 are seeded. Therefore, when the cell culture solution 31 flows into the second groove portion 4a, the gas 32 does not enter the culture chamber 1a.

[0056] When the liquid supply is continued further in the first state 50, the gas 32 enters the second liquid supply tube 4b via the second groove portion 4a, as shown in FIG.

[0057] 13, the cell culture solution 31 flows into the culture solution reservoir 7. That is, the gas 32 contained in the first liquid-supply tube 3b is discharged into the culture solution reservoir 7 through the first groove 3a, the second groove 4a, and the second liquid-supply tube 4b. This removes the gas 32 from the first liquid-supply tube 3b.

[0058] In this embodiment, as shown in FIG. 14, the control unit 6 is configured to control the flow path switching unit 5 to switch from the first state 50 to the second state 51 after the gas 32 contained in the first culture solution flow path 3 is removed.

[0059] After the control unit 6 switches from the first state 50 to the second state 51, the liquid supply from the liquid supply unit 2 is resumed, causing the cell culture solution 31 to flow from the first culture solution flow path 3 (the first liquid supply tube 3b and the first groove 3a) into the culture chamber 1a. As the liquid supply continues, the cell culture solution 31 in the culture chamber 1a is discharged into the culture solution reservoir 7 via the second culture solution flow path 9 (the third groove 9a and the third liquid supply tube 9b), the flow path switching unit 5, and the third discharge flow path 10c. This allows the cell culture solution 31 in the culture chamber 1a to be replaced.

[0060] (Cell culture treatment) Next, the culture process of the cells 30 by the cell culture system 100 will be described with reference to FIG.

[0061] In step 101, cells 30 are seeded in the culture chamber 1a. The process of step 101 may be performed by an operator, or may be performed by the control unit 6 controlling the micropipette 20.

[0062] In step 102, the control unit 6 determines whether a signal to start removing the gas 32 from the first culture solution flow path 3 has been input. If a signal to start removing the gas 32 from the first culture solution flow path 3 has not been input, the control unit 6 repeats the process of step 101. If a signal to start removing the gas 32 from the first culture solution flow path 3 has been input, the process proceeds to step 103. The signal to remove the gas 32 is input by an operator's operation or after a predetermined time has passed since the cells 30 were seeded. The predetermined time until the signal to remove the gas 32 is input is the time required for the cells 30 to adhere to the bottom surface 1b of the culture chamber 1a after being seeded, and is set in advance depending on the type of cells 30 to be seeded and stored in the memory unit 8.

[0063] In step 103, the control unit 6 determines whether or not it is in the first state 50. If it is not in the first state 50, the process proceeds to step 104. If it is in the first state 50, the process proceeds to step 105.

[0064] In step 104 , the control unit 6 controls the flow path switching unit 5 to switch to the first state 50 .

[0065] In step 105, the control unit 6 removes the gas 32 in the first culture solution flow path 3 by sending the cell culture solution 31 through a branch flow path 4 branched from the first culture solution flow path 3 connecting the culture chamber 1a and the liquid delivery unit 2 that delivers the cell culture solution 31 to the culture chamber 1a, in a first state 50 in which the cell culture solution 31 is discharged.

[0066] In step 106, the control unit 6 determines whether a predetermined time has elapsed since the start of the supply of the cell culture solution 31. If the predetermined time has not elapsed, the process of step 106 is repeated. If the predetermined time has elapsed, the process proceeds to step 107. The predetermined time is the time it takes for the gas 32 in the first culture solution flow path 3 to be discharged into the culture solution reservoir 7 via the branch flow path 4. The predetermined time is set in advance based on the diameter of the first culture solution flow path 3, the length of the first culture solution flow path 3, the diameter of the branch flow path 4, the length of the branch flow path 4, the diameter of the third discharge flow path 10c, and the length of the third discharge flow path 10c. The predetermined time for determining whether the gas 32 has been removed is stored in the memory unit 8.

[0067] In step 107, the control unit 6 switches from the first state 50 to a second state 51 in which the cell culture solution 31 enters the culture chamber 1a.

[0068] In step 108, the control unit 6 controls the fluid delivery unit 2 to deliver the cell culture fluid 31 in the second state 51. Then, the process ends.

[0069] (Effects of this embodiment) In this embodiment, the following effects can be obtained.

[0070] In this embodiment, as described above, the cell culture system 100 comprises a cell culture device 1 having a culture chamber 1a capable of accommodating cells 30 and cell culture solution 31, a fluid delivery unit 2 that delivers cell culture solution 31 to the culture chamber 1a, a first culture solution flow path 3 that connects the culture chamber 1a to the fluid delivery unit 2, a branch flow path 4 that branches off from the first culture solution flow path 3 and removes gas 32 from the first culture solution flow path 3, and a flow path switching unit 5 that switches between a first state 50 in which the cell culture solution 31 is discharged through the branch flow path 4 and a second state 51 in which the cell culture solution 31 enters the culture chamber 1a. As a result, even if gas 32 gets mixed into the first culture solution flow path 3 (first liquid supply tube 3b) when connecting the liquid supply unit 2 and the first culture solution flow path 3 (first liquid supply tube 3b), the gas 32 in the first culture solution flow path 3 can be removed via the branch flow path 4 (second groove portion 4a and second liquid supply tube 4b) by discharging the cell culture solution 31 from the branch flow path 4 (second groove portion 4a and second liquid supply tube 4b) in the first state 50. As a result, unlike a configuration in which the first culture solution flow path 3 (first liquid supply tube 3b) and the cell culture device 1 are connected in a state in which the first culture solution flow path 3 (first liquid supply tube 3b) is filled up to its end with cell culture solution 31, it is possible to provide a cell culture system 100 that can prevent gas 32 in the first liquid supply tube 3b from getting into the cell culture device 1 even if gas 32 gets mixed into the first liquid supply tube 3b when connecting the liquid supply unit 2 and the cell culture device 1.

[0071] Furthermore, in this embodiment, as described above, the first culture solution flow path 3 (first groove portion 3a) is provided inside the cell culture device 1, the cell culture device 1 has the second culture solution flow path 9 (third groove portion 9a) that connects to the culture chamber 1a at a position different from the first culture solution flow path 3 (first groove portion 3a), and the branch flow path 4 (second groove portion 4a) is provided branching off from the first culture solution flow path 3 (first groove portion 3a) inside the cell culture device 1. This allows the branch flow path 4 (second groove portion 4a) to branch off from a position closer to the culture chamber 1a, compared to a configuration in which the branch flow path 4 (second fluid supply tube 4b) branches off from the first culture solution flow path 3 (first fluid supply tube 3b) outside the cell culture device 1. As a result, for example, when cells 30 are seeded into the culture chamber 1a via the first culture medium flow path 3, an increase in the amount of cells 30 entering the branch flow path 4 can be suppressed compared to a configuration in which the branch flow path 4 (second liquid supply tube 4b) branches off from the first culture medium flow path 3 (first liquid supply tube 3b) outside the cell culture device 1.

[0072] Furthermore, in this embodiment, as described above, the cell culture device 1 further includes a cell seeding channel 11 connected to the first culture solution channel 3 (first groove portion 3a) for seeding cells 30. The branch portion 3d of the first culture solution channel 3 is located closer to the culture chamber 1a than the connection portion 3e of the cell seeding channel 11 in the first culture solution channel 3 (first groove portion 3a) in the cell culture device 1. This allows gas 32 present at the branch portion 3d to be discharged into the culture chamber 1a when cells 30 are seeded from the cell seeding channel 11. This allows gas 32 present at the position between the branch portion 3d and the culture chamber 1a in the first culture solution channel 3 (first groove portion 3a) to be removed. As a result, intrusion of gas 32 into the culture chamber 1a can be more effectively prevented compared to a configuration in which the connection portion 3e of the cell seeding channel 11 is located closer to the culture chamber 1a than the branch portion 3d.

[0073] In the present embodiment, as described above, the flow path switching unit 5 is connected to the branch flow path 4 (second liquid supply tube 4b) and the second culture solution flow path 9 (third liquid supply tube 9b), and is configured to switch between the first state 50 and the second state 51 by blocking the end of the branch flow path 4 (second liquid supply tube 4b) or the end of the second culture solution flow path 9 (third liquid supply tube 9b). In this manner, in the first state 50, the end of the second culture solution flow path 9 (third liquid supply tube 9b) is blocked, thereby preventing the gas 32 from entering the culture chamber 1a when the gas 32 is removed from the first culture solution flow path 3. In the second state 51, the end of the branch flow path 4 (second liquid supply tube 4b) is blocked, thereby preventing the cell culture solution 31 from entering the branch flow path 4 when the cell culture solution 31 is supplied into the culture chamber 1a. As a result, in the first state 50, it is possible to effectively prevent the gas 32 in the first culture solution flow path 3 from entering the culture chamber 1a, and in the second state 51, it is possible to effectively prevent the cell culture solution 31 from being discharged from the branch flow path 4.

[0074] Furthermore, in this embodiment, as described above, the flow path switching unit 5 includes a switching valve 5a that switches a discharge flow path for discharging the cell culture solution 31, which has been delivered from the fluid delivery unit 2, from the cell culture device 1, and the discharge flow path includes a first discharge flow path 10a formed by the first culture solution flow path 3 (the first fluid delivery tube 3b and a part of the first groove portion 3a) and the branch flow path 4, and a second discharge flow path 10b formed by the first culture solution flow path 3, the culture chamber 1a, and the second culture solution flow path 9. This allows easy and reliable switching between the first state 50 and the second state 51 by switching between the first discharge flow path 10a and the second discharge flow path 10b using the switching valve 5a.

[0075] Furthermore, as described above, this embodiment further includes a control unit 6 that controls switching between the first state 50 and the second state 51 by the flow path switching unit 5, and the control unit 6 is configured to control switching from the first state 50 to the second state 51 by the flow path switching unit 5 after the gas 32 contained in the first culture solution flow path 3 is removed. In this way, switching between the first state 50 and the second state 51 by the control unit 6 can reduce the burden on the operator compared to a configuration in which the operator manually operates the flow path switching unit 5 to switch between the first state 50 and the second state 51 after the gas 32 is removed from the first culture solution flow path 3.

[0076] Furthermore, in this embodiment, as described above, the cell culture method is a cell culture method for culturing cells 30 using a cell culture device 1 having a culture chamber 1a capable of accommodating cells 30 and cell culture solution 31, and includes the steps of seeding cells 30 in the culture chamber 1a, removing gas 32 in the first culture solution flow path 3 by sending cell culture solution 31 through a branch flow path 4 branched from a first culture solution flow path 3 connecting the culture chamber 1a and a liquid delivery section 2 that delivers cell culture solution 31 to the culture chamber 1a in a first state 50 in which the cell culture solution 31 is discharged, switching from the first state 50 to a second state 51 in which the cell culture solution 31 enters the culture chamber 1a, and delivering the cell culture solution 31 in the second state 51. As a result, unlike the configuration in which the first culture solution flow path 3 (first liquid supply tube 3b) is connected to the cell culture device 1 when the first culture solution flow path 3 (first liquid supply tube 3b) is filled with cell culture solution 31 up to the end, as in the cell culture system 100 described above, a cell culture method can be provided that can prevent gas 32 in the first liquid supply tube 3b from entering the cell culture device 1 even if gas 32 gets mixed into the first liquid supply tube 3b when connecting the liquid supply section 2 to the cell culture device 1.

[0077] [Variations] The embodiments disclosed herein should be considered to be illustrative and not restrictive in all respects. The scope of the present invention is defined by the claims rather than the above description of the embodiments, and further includes all modifications (variations) within the meaning and scope of the claims.

[0078] (First Modification) For example, in the above embodiment, an example of a configuration in which the flow path switching unit 5 is connected to the branch flow path 4 (second fluid supply tube 4b) and the second culture solution flow path 9 (third fluid supply tube 9b) has been shown, but the present invention is not limited to this. For example, as shown in Fig. 17, the flow path switching unit 5 may be provided at the branch point 3d between the first culture solution flow path 3 (first fluid supply tube 3b) and the branch flow path 4 (second fluid supply tube 4b). Furthermore, when the flow path switching unit 5 is provided at the branch point 3d between the first culture solution flow path 3 (first fluid supply tube 3b) and the branch flow path 4 (second fluid supply tube 4b), the cell seeding flow path 11 may be connected to the first culture solution flow path 3 (first fluid supply tube 3b).

[0079] When the flow path switching unit 5 is provided at the branch portion 3d as in the first modified example shown in Fig. 17, the switching valve 5a may include, for example, a three-way valve. When the switching valve 5a is a three-way valve, the state in which the first culture solution flow path 3 (first liquid supply tube 3b) and the branch flow path 4 (second liquid supply tube 4b) are connected is the first state 50, as shown in Fig. 17. Furthermore, the state in which the first culture solution flow path 3 (first liquid supply tube 3b) is connected to the culture chamber 1a is the second state 51, as shown in Fig. 18.

[0080] In the first modified example, the control unit 6 is configured to remove the gas 32 in the first culture solution flow path 3 (first solution supply tube 3b) in the first state 50, and then switch to the second state 51 to control the supply of the cell culture solution 31 into the culture chamber 1a. The other configurations of the first modified example are the same as those of the above embodiment.

[0081] In the first modified example, as described above, the flow path switching unit 5 is provided at the branching portion 3d between the first culture solution flow path 3 (first liquid supply tube 3b) and the branch flow path 4 (second liquid supply tube 4b). This makes it possible to switch between the first state 50 and the second state 51 even when the flow path switching unit 5 is provided at the branching portion 3d between the first culture solution flow path 3 (first liquid supply tube 3b) and the branch flow path 4 (second liquid supply tube 4b), thereby preventing the gas 32 in the first culture solution flow path 3 from entering the culture chamber 1a. As a result, the degree of freedom in arranging the flow path switching unit 5 can be improved. Other effects of the first modified example are similar to those of the above embodiment.

[0082] (Second Modification) In the above embodiment, an example of a configuration in which the cell culture solution 31 is discharged from the cell culture device 1 to the culture solution reservoir 7 via the second culture solution flow path 9 (third liquid supply tube 9b) has been shown, but the present invention is not limited to this. For example, as shown in Fig. 19, a liquid supply tube 13 may be provided that connects the liquid supply unit 2 and the culture solution reservoir 7 and returns the cell culture solution 31 stored in the culture solution reservoir 7 to the liquid supply unit 2.

[0083] As shown in FIG. 19 , in the second modified example, one end of the fluid supply pipe 13 is connected to the culture medium storage section 7, and the other end is connected to the fluid supply section 2. In the second modified example, the fluid supply section 2 is configured to obtain the cell culture medium 31 from the culture medium storage section 7 via the fluid supply pipe 13. That is, the fluid supply section 2 according to the second modified example circulates the cell culture medium 31 in the cell culture system 100. The fluid supply section 2 according to the second modified example includes, for example, a tube pump. The other configurations of the second modified example are the same as those of the above embodiment.

[0084] As described above, the second modified example further includes a culture medium reservoir 7 connected to the flow path switching unit 5 and configured to store the cell culture medium 31 discharged to the outside of the cell culture device 1, and a fluid delivery tube 13 connecting the fluid delivery unit 2 to the culture medium reservoir 7 and returning the cell culture medium 31 stored in the culture medium reservoir 7 to the fluid delivery unit 2. The fluid delivery unit 2 is configured to retrieve the cell culture medium 31 from the culture medium reservoir 7 via the fluid delivery tube 13. This allows the cell culture medium 31 to be circulated in the cell culture system 100 by delivering the cell culture medium 31 retrieved from the culture medium reservoir 7 to the first culture medium flow path 3. Furthermore, since the cell culture medium 31 can be circulated, for example, by connecting multiple cell culture devices 1 seeded with different types of cells, metabolites of the cells 30 can be circulated. As a result, by connecting multiple cell culture devices 1 seeded with different types of cells, a simulated internal body environment can be reproduced. Other advantages of the second modified example are similar to those of the above embodiment.

[0085] (Third Modification) Furthermore, in the above embodiment, an example of a configuration in which the cell culture device 1 has the cell seeding channel 11 and the cells 30 are seeded through the cell seeding channel 11 has been described, but the present invention is not limited to this. For example, the cell culture device 1 does not have to have the cell seeding channel 11. When the cell culture device 1 does not have the cell seeding channel 11, the culture chamber 1a only needs to have an opening 1d in the upper surface portion 1c, as in a third modified example shown in FIG.

[0086] In the third modified example, the culture chamber 1a has an opening 1d in the upper surface portion 1c. The cell culture device 1 also has a lid 1e that covers the opening 1d of the culture chamber 1a. When seeding cells 30 into the culture chamber 1a, the lid 1e is removed and the cells are seeded with the opening 1d open. Thereafter, the lid 1e is closed and the cells 30 are cultured in a thermostatic bath. The other configurations of the third modified example are the same as those of the above embodiment.

[0087] In the third modified example, as described above, the culture chamber 1a has an opening 1d in the upper surface portion 1c, and the cell culture device 1 further includes a lid 1e that covers the opening 1d of the culture chamber 1a. With this configuration, the cells 30 can be easily seeded by opening the opening 1d and seeding the cells 30 into the culture chamber 1a through the opening 1d. Furthermore, the cells 30 can be easily recovered by recovering them from the culture chamber 1a through the opening 1d. As a result, the convenience for the operator can be improved. Other effects of the third modified example are the same as those of the above embodiment.

[0088] (Fourth Modification) Furthermore, in the above embodiment, an example of a configuration in which the control unit 6 determines whether the gas 32 in the first culture solution flow path 3 has been removed by determining whether a predetermined time has elapsed has been described, but the present invention is not limited to this. For example, as in a fourth modified example shown in FIG. 21 , the cell culture system 100 may further include a detection unit 14 that detects the gas 32 in the second liquid supply tube 4b. The control unit 6 may be configured to determine that the gas 32 in the first culture solution flow path 3 has been removed by the detection unit 14 having caused the gas 32 to pass through the second liquid supply tube 4b. The detection unit 14 may be configured to detect the gas 32 in the second liquid supply tube 4b by acquiring an image of the second liquid supply tube 4b. The detection unit 14 may include, for example, an image sensor and an optical system such as a lens.

[0089] In the fourth modified example, as described above, the control unit 6 is configured to determine whether or not the gas 32 has been removed from the first culture solution flow path 3 using the detection unit 14. With this configuration, the first state 50 can be quickly switched to the second state 51 after the gas 32 has been removed from the first culture solution flow path 3. As a result, the amount of cell culture solution 31 discarded can be reduced compared to a configuration in which the first state 50 is switched to the second state 51 after a predetermined time has elapsed. Other effects of the fourth modified example are similar to those of the above embodiment.

[0090] (Other variations) Furthermore, in the above embodiment, an example of a configuration in which the cell culture system 100 includes the control unit 6 has been described, but the present invention is not limited to this. For example, the cell culture system 100 does not have to include the control unit 6. When the cell culture system 100 does not include the control unit 6, after the gas 32 in the first culture solution flow path 3 is removed, the operator can switch between the first state 50 and the second state 51 by operating the flow path switching unit 5. However, when the operator switches between the first state 50 and the second state 51, the burden on the operator increases. Therefore, it is preferable that the cell culture system 100 includes the control unit 6.

[0091] Furthermore, in the above embodiment, an example of a configuration in which the branch portion 3d is located closer to the culture chamber 1a than the connection portion 3e has been described, but the present invention is not limited to this. The branch portion 3d may be located farther from the culture chamber 1a than the connection portion 3e. However, if the branch portion 3d is located farther from the culture chamber 1a than the connection portion 3e, gas 32 may remain between the branch portion 3d and the connection portion 3e after cells 30 are seeded through the cell seeding channel 11. In this case, when the cell culture solution 31 is delivered in the second state 51, the gas 32 may enter the culture chamber 1a. Therefore, it is preferable to locate the branch portion 3d closer to the culture chamber 1a than the connection portion 3e.

[0092] Furthermore, in the above embodiment, the branch channel 4 includes the second groove portion 4a and the second liquid supply tube 4b, and the second groove portion 4a branches off from the first groove portion 3a inside the cell culture device 1. However, the present invention is not limited to this. For example, the branch channel 4 may include only the second liquid supply tube 4b and be configured to branch off from the first culture solution channel 3 (the first liquid supply tube 3b) outside the cell culture device 1. However, in a configuration in which the branch channel 4 (the second liquid supply tube 4b) branches off from the first culture solution channel 3 (the first liquid supply tube 3b) outside the cell culture device 1, the connection portion 3e of the cell seeding channel 11 must also be provided outside the cell culture device 1 in order to remove the gas 32 inside the first culture solution channel 3. In this case, the distance between the branch portion 3d and the culture chamber 1a becomes larger. Therefore, compared to a configuration in which the branch channel 4 branches off inside the cell culture device 1, the amount of cell culture solution 31 flowing into the branch channel 4 when the cells 30 are seeded increases. Therefore, it is preferable that the branch channel 4 be provided inside the cell culture device 1.

[0093] In the above embodiment, the liquid delivery unit 2 is a syringe pump, but the present invention is not limited to this. For example, the liquid delivery unit 2 may be configured with a gravity pump that uses gravity, a droplet surface tension pump that is driven by the surface tension of droplets, an osmotic flow pump that is driven by osmotic pressure, or a negative pressure pump that draws the cell culture solution 31 by negative pressure. The liquid delivery unit 2 may be any pump as long as it is capable of delivering the cell culture solution 31.

[0094] [Aspect] It will be appreciated by those skilled in the art that the exemplary embodiments described above are examples of the following aspects.

[0095] (Item 1) a cell culture device having a culture chamber capable of containing cells and a cell culture medium; a fluid delivery unit that delivers the cell culture solution to the culture chamber; a first culture solution flow path connecting the culture chamber and the solution delivery unit; a branch flow path branched from the first culture solution flow path for removing gas from the first culture solution flow path; A cell culture system comprising: a flow path switching unit that switches between a first state in which the cell culture medium is discharged through the branch flow path and a second state in which the cell culture medium enters the culture chamber.

[0096] (Item 2) the first culture solution flow path is provided in the cell culture device, the cell culture device has a second culture medium flow path connected to the culture chamber at a position different from the first culture medium flow path; Item 2. The cell culture system according to item 1, wherein the branch channel is provided by branching off from the first culture medium channel in the cell culture device.

[0097] (Item 3) the cell culture device further includes a cell seeding channel connected to the first culture solution channel for seeding cells; 3. The cell culture system according to item 2, wherein the branching portion of the first culture solution flow path is provided in the first culture solution flow path in the cell culture device at a position closer to the culture chamber than the connection portion of the cell seeding flow path.

[0098] (Item 4) Item 4. The cell culture system according to item 3, wherein the flow path switching unit is connected to the branch flow path and the second culture solution flow path, and is configured to switch between the first state and the second state by blocking an end of the branch flow path or an end of the second culture solution flow path.

[0099] (Item 5) the flow path switching unit includes a switching valve for switching a discharge flow path that discharges the cell culture solution delivered from the solution delivery unit from the cell culture device, 5. The cell culture system according to item 4, wherein the discharge flow path includes a first discharge flow path formed by the first culture solution flow path and the branch flow path, and a second discharge flow path formed by the first culture solution flow path, the culture chamber, and the second culture solution flow path.

[0100] (Item 6) a control unit that controls switching between the first state and the second state by the flow path switching unit, 6. The cell culture system according to item 4 or 5, wherein the control unit is configured to control the flow path switching unit to switch from the first state to the second state after gas contained in the first culture solution flow path is removed.

[0101] (Item 7) Item 2. The cell culture system according to item 1, wherein the flow path switching unit is provided at a branch point between the first culture solution flow path and the branch flow path.

[0102] (Item 8) a culture medium storage unit connected to the flow path switching unit and configured to store the cell culture medium to be discharged to the outside of the cell culture device; a fluid transfer pipe that connects the fluid transfer unit and the culture medium storage unit and returns the cell culture medium stored in the culture medium storage unit to the fluid transfer unit; 8. The cell culture system according to any one of items 1 to 7, wherein the fluid delivery unit is configured to obtain the cell culture fluid from the culture fluid reservoir unit via the fluid delivery pipe.

[0103] (Item 9) the culture chamber has an opening in an upper surface portion, 9. The cell culture system according to any one of items 1 to 8, wherein the cell culture device further comprises a lid portion that covers the opening of the culture chamber.

[0104] (Item 10) A cell culture method for culturing cells using a cell culture device having a culture chamber capable of accommodating cells and a cell culture medium, comprising: seeding the culture chamber with cells; a step of removing gas from within the first culture solution flow path by sending the cell culture solution through a branch flow path branched from a first culture solution flow path connecting the culture chamber and a solution sending unit that sends the cell culture solution to the culture chamber in a first state in which the cell culture solution is discharged; switching from the first state to a second state in which the cell culture medium enters the culture chamber; and delivering the cell culture medium in the second state. [Explanation of symbols]

[0105] 1. Cell culture device 2. Liquid delivery section 3 First culture solution flow path 3a 1st groove (1st culture solution flow path) 3b First liquid supply tube (first culture medium flow path) 4 Branching channels 4a Second groove portion (branch flow path) 4b Second liquid supply tube (branch flow path) 5 Flow path switching section 5a Switching valve 6 Control Unit 7. Culture medium reservoir 9 Second culture medium flow path 9a Third groove (second culture fluid flow path) 9b Third liquid supply tube (second culture medium flow path) 10a First discharge flow path (discharge flow path) 10b Second discharge flow path (discharge flow path) 11. Cell seeding channel 30 cells 31 Cell culture medium 32 Gas 50 First state 51 Second State 100 Cell Culture Systems

Claims

1. a cell culture device having a culture chamber capable of containing cells and a cell culture medium; a fluid delivery unit provided outside the cell culture device and configured to deliver the cell culture solution to the culture chamber; a first culture medium flow path, a portion of which is provided within the cell culture device and which connects the culture chamber and the fluid delivery unit; a branch flow path branched from the first culture solution flow path for removing gas from the first culture solution flow path; a flow path switching unit that switches between a first state in which the cell culture solution is discharged through the branch flow path and a second state in which the cell culture solution enters the culture chamber; a cell seeding flow path connected to the first culture medium flow path and through which a cell suspension flows; a control unit that controls switching between the first state and the second state by the flow path switching unit, a branched portion between the first culture solution flow path and the branched flow path is provided in the first culture solution flow path in the cell culture device at a position closer to the culture chamber than a connection portion of the cell seeding flow path, The control unit switches from the first state to the second state using the flow path switching unit when a predetermined time has elapsed since the cell culture solution started to be sent into the first culture solution flow path in the first state, and the cell suspension has filled the portion of the culture chamber closer to the branching portion.

2. 2. The cell culture system according to claim 1, wherein the flow path switching unit is connected to a second culture medium flow path and the branch flow path that connect to the culture chamber at a position different from the first culture medium flow path, and is configured to switch between the first state and the second state by blocking an end of the branch flow path or an end of the second culture medium flow path.

3. the flow path switching unit includes a switching valve for switching a discharge flow path that discharges the cell culture solution delivered from the solution delivery unit from the cell culture device, 3. The cell culture system according to claim 2, wherein the discharge flow path includes a first discharge flow path formed by the first culture solution flow path and the branch flow path, and a second discharge flow path formed by the first culture solution flow path, the culture chamber, and the second culture solution flow path.

4. a culture medium storage unit connected to the flow path switching unit and configured to store the cell culture medium to be discharged to the outside of the cell culture device; a fluid transfer pipe that connects the fluid transfer unit and the culture medium storage unit and returns the cell culture medium stored in the culture medium storage unit to the fluid transfer unit; The cell culture system according to any one of claims 1 to 3, wherein the fluid delivery unit is configured to acquire the cell culture fluid from the culture fluid storage unit via the fluid delivery pipe.

5. the culture chamber has an opening in an upper surface portion, The cell culture system according to any one of claims 1 to 4, wherein the cell culture device further comprises a lid portion that covers the opening of the culture chamber.

6. A cell culture method for culturing cells using a cell culture device having a culture chamber capable of accommodating cells and a cell culture medium, comprising: seeding the culture chamber with cells; a step of connecting the culture chamber with a fluid delivery unit that is provided outside the cell culture device and that delivers the cell culture solution to the culture chamber, and delivering the cell culture solution through a branched fluid path that is provided in the cell culture device and is partly branched from a first culture solution path provided inside the cell culture device, in a first state in which the cell culture solution is discharged, thereby delivering the cell culture solution; switching from the first state to a second state in which the cell culture medium enters the culture chamber; and delivering the cell culture medium in the second state, a branching portion between the first culture solution flow path and the branching flow path is provided in the first culture solution flow path in the cell culture device at a position closer to the culture chamber than a connecting portion of a cell seeding flow path connected to the first culture solution flow path and through which a cell suspension flows, In the step of switching from the first state to the second state, a predetermined time has elapsed since the cell culture solution began to be supplied into the first culture solution flow path in the first state, and the cell suspension has filled the portion closer to the culture chamber than the branching portion, and the cell culture method is switched from the first state to the second state.

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