Cell culture device

Abstract

To provide a cell culture apparatus in which an excessive length of a liquid tube connected to a cell culture vessel is appropriately set.SOLUTION: A cell culture apparatus comprises: a base; a rotary table 32 which is rotatably attached to the base about a first axis X; a vessel holding member 33 which is rotatably attached to the rotary table about a second axis Y and holds a cell culture vessel 2; and a drive mechanism 30 which rotationally drives the rotary table about the first axis X and rotationally drives the vessel holding member about the second axis Y. A liquid supply / drain port 25 to which a liquid supply / drain tube 50 is connected is arranged on the outer peripheral surface of the cell culture vessel, a first prescribed part 51 which is apart from a connection part 50a connected to the liquid supply / drain port by a prescribed length in the liquid supply / drain tube is positioned at the vessel holding member by a first positioning member 61, a second prescribed part 52 which is apart from the connection part further than the first prescribed part in the liquid supply / drain tube is positioned at the rotary table by a second positioning member 62, and the first positioning member is rotatably attached to the vessel holding member.SELECTED DRAWING: Figure 4

Description

[Technical field]

[0001] The present invention relates to a cell culture device suitable for appropriately carrying out large-scale culture. [Background technology]

[0002] In cell culture techniques that use cell culture vessels for culture, a cell culture device is used that performs handling operations such as rotating the cell culture vessel in order to introduce culture medium in which cells are seeded into the cell culture vessel or to recover the culture medium from the cell culture vessel (see, for example, Patent Document 1).

[0003] The cell culture device of Patent Document 1 includes a drive mechanism for changing the posture of a cell culture vessel. The drive mechanism includes a base, a turntable rotatably attached to the base about a first axis X, a vessel holding member rotatably attached to the turntable about a second axis Y, a first drive unit that drives the turntable to rotate about the first axis X, a second drive unit that drives the vessel holding member to rotate about the second axis Y, and a controller that controls these drive units.

[0004] Therefore, by controlling the first drive unit and the second drive unit, the turntable and / or container holding member can be rotated to supply culture medium from the supply / discharge port of the cell culture container or recover culture medium from the supply / discharge port of the cell culture container. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2020-103232 A Summary of the Invention [Problem to be solved by the invention]

[0006] In the cell culture device of Patent Document 1, a supply / discharge tube for supplying and discharging a culture medium is connected to a supply / discharge port of a cell culture container, and the turntable and / or container holding member are rotated while the supply / discharge tube is connected to the supply / discharge port. In order to prevent the supply / discharge tube from getting caught on the surroundings when the turntable and / or container holding member rotates, multiple parts of the supply / discharge tube that are separated from each other are positioned on the turntable or container holding member. In this case, an excess part is formed between the part of the supply / discharge tube positioned on the turntable and the part positioned on the container holding member so that the rotation of the turntable and / or container holding member is not restricted.

[0007] However, if the excess length of the liquid supply and drainage tube is made too long, there is a problem that the liquid supply and drainage tube gets caught on the surroundings when the turntable and / or container holding member rotates, causing the liquid supply and drainage tube to tear or come off the liquid supply and drainage port, making it impossible to continue the culture. Also, if the excess length of the liquid supply and drainage tube is made too short, there is a problem that the liquid supply and drainage tube is pulled when the turntable and / or container holding member rotates, causing the liquid supply and drainage tube to come off the liquid supply and drainage port or to bend and become blocked, making it impossible to continue the culture.

[0008] The present invention has been made in view of the above-mentioned problems, and has an object to provide a cell culture device in which the extra length of a liquid tube connected to a cell culture vessel is appropriately set. [Means for solving the problem]

[0009] In order to achieve the above object, the present invention takes the following measures.

[0010] That is, the cell culture device of the present invention includes a base, a turntable rotatably attached to the base about a first axis, a container holding member rotatably attached to the turntable about a second axis and holding a cell culture container, and a drive mechanism for rotatably driving the turntable about the first axis and rotatably driving the container holding member about the second axis, wherein a liquid port to which a liquid tube is connected is disposed on an outer peripheral surface of the cell culture container, a first predetermined portion of the liquid tube that is a predetermined length away from a connection portion connected to the liquid port is positioned on the container holding member by a first positioning member, and a second predetermined portion of the liquid tube that is further away from the connection portion than the first predetermined portion is positioned on the turntable by a second positioning member, and the first positioning member is rotatably attached to the container holding member. When the first positioning member that positions the first predetermined portion rotates relative to the container holding member, the orientation of the liquid tube on the upstream side of the first predetermined portion and the orientation of the liquid tube on the downstream side of the first predetermined portion change. It is characterized by:

[0011] With this, by rotatably mounting the first positioning member that positions the first predetermined portion of the liquid tube to the container holding member, it is possible to eliminate unnecessary slack formed between the first predetermined portion and the second predetermined portion of the liquid tube. Therefore, when supplying a reagent into the cell culture vessel or draining a reagent from the cell culture vessel, even if the turntable and / or the container holding member are rotated while the liquid tube is connected to the liquid port, it is possible to prevent the slack length of the liquid tube connected to the liquid port from becoming too long so that culture can be continued appropriately.

[0012] The cell culture device of the present invention comprises a base, a turntable rotatably attached to the base about a first axis, a container holding member rotatably attached to the turntable about a second axis and holding a cell culture container, and a drive mechanism for rotationally driving the turntable about the first axis and the container holding member about the second axis, wherein a liquid port to which a liquid tube is connected is arranged on the outer peripheral surface of the cell culture container, a first predetermined portion of the liquid tube that is a predetermined length away from a connection portion connected to the liquid port is positioned on the container holding member by a first positioning member, and a second predetermined portion of the liquid tube that is further away from the connection portion than the first predetermined portion is positioned on the turntable by a second positioning member, and the first positioning member positions the first predetermined portion on the container holding member at a position that is not on the second axis and is rotatably attached to the container holding member.

[0013] With this, by rotatably mounting the first positioning member that positions the first predetermined portion of the liquid tube to the container holding member, it is possible to eliminate unnecessary slack formed between the first predetermined portion and the second predetermined portion of the liquid tube. Therefore, when supplying a reagent into the cell culture vessel or draining a reagent from the cell culture vessel, even if the turntable and / or the container holding member are rotated while the liquid tube is connected to the liquid port, it is possible to prevent the slack length of the liquid tube connected to the liquid port from becoming too long so that culture can be continued appropriately.

[0014] The cell culture device of the present invention comprises a base, a turntable rotatably attached to the base about a first axis, a container holding member rotatably attached to the turntable about a second axis and holding a cell culture container, and a drive mechanism for rotatably driving the turntable about the first axis and rotatably driving the container holding member about the second axis, wherein a liquid port to which a liquid tube is connected is arranged on the outer peripheral surface of the cell culture container, a first predetermined portion of the liquid tube that is a predetermined length away from a connection portion connected to the liquid port is positioned on the container holding member by a first positioning member, a second predetermined portion of the liquid tube that is farther away from the connection portion than the first predetermined portion is positioned on the turntable by a second positioning member, and a third predetermined portion of the liquid tube that is closer to the connection portion than the first predetermined portion is positioned on the container holding member by a third positioning member, the first positioning member is rotatably attached to the container holding member, and the first predetermined portion and the third predetermined portion are positioned on the same plane with respect to the container holding member.

[0015] This makes it easier for the direction of the first predetermined portion of the liquid tube, which is rotatably positioned relative to the container holding member by the first positioning member, to change when the turntable and / or the container holding member is rotated while the liquid tube is connected to the liquid port. This makes it easier to eliminate excess length formed between the first predetermined portion and the second predetermined portion of the liquid tube. Also,The orientation of the first predetermined portion of the liquid tube, which is rotatably positioned with respect to the container holding member by the first positioning member, becomes easier to change, and therefore, the excess length formed between the first predetermined portion and the second predetermined portion of the liquid tube can be further eliminated.

[0016] In the cell culture device of the present invention, the driving mechanism is characterized in that, during a drainage operation to drain reagent from the cell culture container, it drives the rotating table and the container holding member to rotate so that the reagent flowing out from the liquid port into the liquid tube flows through the liquid tube in a direction away from the liquid port due to gravity acting on it.

[0017] This makes it possible to prevent the reagent from remaining in the liquid tube when the reagent is drained from the cell culture vessel. Effect of the Invention

[0018] The present invention can provide a cell culture device in which the extra length of the liquid tube connected to the liquid port is appropriately set. Also, the amount of liquid remaining in the liquid tube can be reduced. [Brief description of the drawings]

[0019] [Figure 1] 1 is a schematic configuration diagram of a cell culture system according to an embodiment of the present invention. [Diagram 2] FIG. 2 is a diagram showing an incubator accommodating a cell culture vessel in the cell culture system of FIG. 1. [Diagram 3] FIG. 2 is a diagram showing a shelf structure of a cell culture vessel. [Figure 4] FIG. 2 is a diagram showing a drive mechanism of the cell culture device. [Diagram 5] FIG. 2 is a diagram showing a drive mechanism of the cell culture device. [Figure 6] FIG. 2 is a diagram showing a drive mechanism of the cell culture device. [Figure 7] 13A to 13C are diagrams showing changes in state of the liquid supply and drainage tube when the container holding member is rotationally driven about the second axis Y. [Figure 8]13A to 13C are diagrams showing changes in state of the liquid supply and drainage tube when the container holding member is rotationally driven about the second axis Y. [Figure 9] FIG. [Figure 10] FIG. 13 is a diagram illustrating a liquid drainage operation. [Figure 11] 11A and 11B are diagrams illustrating a first liquid drainage posture in a liquid drainage operation. [Figure 12] 13A and 13B are diagrams illustrating a second liquid drainage posture in the liquid drainage operation. [Figure 13] 13 is a diagram illustrating a third liquid drainage posture in the liquid drainage operation. FIG. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

[0021] 1 is a system diagram showing a simplified view of the cell culture system of this embodiment. This cell culture system includes a liquid storage section 1, such as a refrigerator, a cell culture vessel 2, and a cell recovery section 3 that recovers cells cultured in the cell culture vessel 2. The liquid storage section 1, the cell culture vessel 2, and the cell recovery section 3 are connected at appropriate locations with sterile connectors (not shown), and once operation is started after assembly, the system is kept as a closed line until operation is stopped, preventing the intrusion of bacteria from the outside.

[0022] The liquid reservoir 1 stores liquids such as culture medium and various reagents to be supplied to the cell culture vessel 2, and the inside is kept at a temperature required to keep the culture medium and various reagents cool, for example, about 4°C.

[0023] The cell culture vessel 2 has a shelf structure in which cells are cultured in a medium spread out on a flat surface, and is placed in a temperature environment of about 37° C. to promote cell division.

[0024] The cell recovery unit 3 recovers cells in the liquid removed after the completion of culture in the cell culture vessel 2. Between the cell culture vessel 2 and the cell recovery unit 3, an intermediate processing unit 4 is interposed, which includes an analysis unit for analyzing the state of the cells as necessary and an adjustment unit for adjusting the dilution degree of the suspension containing the cells. All the lines in this intermediate processing unit 4 are also composed of closed lines.

[0025] In such a closed line, a system is constructed which automatically performs the following steps without human intervention: preparing liquids such as culture medium and reagents in liquid storage section 1, transferring those liquids from liquid storage section 1 to cell culture vessel 2, culturing cells in cell culture vessel 2, removing the cultured cells from cell culture vessel 2 and recovering them in cell recovery section 3, and performing various intermediate processing steps in intermediate processing section 4 on the liquid removed from cell culture vessel 2 before recovery.

[0026] For this purpose, a valve V and a pump P (pump P(a) and pump P(b)) are connected to each line as necessary, and these valves V and pumps P are controlled by control commands from a controller C. Pumps P(a) and P(b) can be used for both pressure delivery and suction.

[0027] In this embodiment, in order to maintain the cell culture vessel 2 at a predetermined temperature without breaking the closed system line, the cell culture device 10 including the cell culture vessel 2 and peripheral components is placed in an incubator 5 as shown in Figures 1 and 2 with the supply and drainage tube 50 that forms part of the closed system line still connected.

[0028] This places spatial restrictions on the cell culture vessel 2. Therefore, in order to enable as much culture as possible, it is necessary to increase the culture rate in the cell culture vessel 2.

[0029] Therefore, in this embodiment, a configuration is adopted in which the cell culture vessel 2 has a multi-tiered shelf structure, and the culture area is increased by the number of shelves 21. Due to the multi-tiered shelf structure, a process is required in which a liquid supplied from the outside is spread across each shelf 21, and after the culture is completed, the liquid is drained from each shelf 21 to the outside. In this case, in order to reduce the number of connected closed system lines as much as possible, a liquid supply and drainage port 25 serving as both a liquid supply port and a liquid drainage port is provided on the housing of the cell culture vessel 2, and the liquid supply and drainage port 25 is common to each shelf 21, and the cell culture vessel 2 is changed in position while maintaining a closed line state in the incubator 5, thereby realizing the supply of liquid to each shelf 21 and the drainage from each shelf 21.

[0030] Specifically, as shown in Fig. 3, the cell culture vessel 2 is configured such that a part of the shelf surface 21b is cut out to provide a communication part 21c at the corner part on one edge 21a side of each shelf 21 so that liquid can flow evenly into each shelf space, and the shelf spaces are partitioned by the shelf surface 21b and the side surface 21d in the parts other than the communication part 21c. At the boundary between the communication part 21c and the shelf surface 21b, a standing wall 21e having a height lower than the side surface 21d is provided so that liquid can be stored when the shelf surface 21b is in a horizontal state and the liquid can be discharged when the shelf surface 21b is tilted or inverted. In other words, the hatched part in Fig. 3(b) indicates the area where liquid can be stored when the shelf surface 21b is in a horizontal state.

[0031] As shown in Figures 4 to 6, the cell culture device 10 includes a drive mechanism 30 for changing the attitude of the cell culture vessel 2. Note that Figure 4 illustrates a liquid supply / drain tube 50 that serves both as a liquid tube for supplying liquid to the cell culture vessel 2 and as a liquid tube for draining liquid from the cell culture vessel 2, but Figures 5 to 6 omit the illustration of the liquid supply / drain tube 50.

[0032] The drive mechanism 30 comprises a base 31, a rotating table 32 rotatably attached to the base 31 about a first axis X, a container holding member 33 rotatably attached to the rotating table 32 about a second axis Y, a first drive unit 34 that drives and rotates the rotating table 32 about the first axis X, a second drive unit 35 that drives and rotates the container holding member 33 about the second axis Y, and a controller C that serves as a control unit for controlling these drive units 34, 35.

[0033] As shown in Fig. 4, a left rotation around the first axis X is considered to be a positive (+) rotation, and a right rotation around the first axis X is considered to be a negative (-) rotation. Similarly, a left rotation around the second axis Y is considered to be a positive (+) rotation, and a right rotation around the second axis Y is considered to be a negative (-) rotation.

[0034] The first drive unit 34 is located in a direction along the first axis X from the cell culture vessel 2, and the second drive unit 35 is disposed near the first drive unit 34 and is connected to the second axis Y via a transmission mechanism 36 using a belt, chain, or the like that transmits power along the first axis X.

[0035] That is, when only the first driving unit 34 is driven, the turntable 32, the container holding member 33, and the cell culture vessel 2 rotate around the first axis X as shown in Fig. 5, and when only the second driving unit 35 is driven, the turntable 32 does not rotate but the container holding member 33 and the cell culture vessel 2 rotate around the second axis Y as shown in Fig. 6, and further, when the first driving unit 34 and the second driving unit 35 are driven simultaneously, the turntable 32, the container holding member 33, and the cell culture vessel 2 rotate around the first axis X as shown in Fig. 5, and further, the container holding member 33 and the cell culture vessel 2 can rotate around the second axis Y relative to the turntable 32 as shown in Fig. 6. The driving mechanism 30 may control the attitude of the cell culture vessel 2 by open control, or may provide a sensor or the like that detects the attitude of the shelf surface 21b to feedback control the attitude of the cell culture vessel 2.

[0036] A liquid supply / drain port 25, which serves as both a liquid supply port and a liquid drain port, is provided on the outer circumferential surface of the cell culture vessel 2. A liquid supply / drain tube 50, which is connected to the liquid storage section 1 and the cell recovery section 3, is connected to the liquid supply / drain port 25. The liquid supply / drain tube 50 is used to supply a reagent into the cell culture vessel 2 and to drain the reagent from within the cell culture vessel 2.

[0037] 4 shows a state where the shelf surface 21b of the cell culture vessel 2 is placed in a horizontal culture position, and in this state, the vessel holding member 33 has a substantially rectangular parallelepiped shape with an open upper surface. A substantially rectangular opening 33b is formed in a front portion 33a of the vessel holding member 33. In the state shown in FIG. 4, the turntable 32 is disposed so as to face the right side surface and rear surface of the vessel holding member 33.

[0038] A first predetermined portion 51 of the liquid supply / drainage tube 50, which is a predetermined length away from the connection portion 50a connected to the liquid supply / drainage port 25 along the liquid tube longitudinal direction, is positioned in the container holding member 33 by a first positioning member 61. The first positioning member 61 that holds the first predetermined portion 51 is disposed near the upper end of the front portion 33a of the container holding member 33 and to the right of the opening 33b.

[0039] A second predetermined portion 52, which is farther away from the connection portion 50a connected to the supply / drain port 25 in the supply / drain tube 50 than the first predetermined portion 51, is positioned on the turntable 32 by a second positioning member 62. The second positioning member 62 is disposed on the turntable 32 at a front portion 32a of a plate-like member that faces the right side portion of the container holding member 33.

[0040] The third predetermined portion 53 of the liquid supply / drainage tube 50, which is closer to the connection portion 50a than the first predetermined portion 51, is positioned on the container holding member 33 by the third positioning member 63. The third positioning member 63 is positioned on the attachment portion 33c that protrudes upward from the left side of the opening 33b at the upper end of the front portion 33a of the container holding member 33. Therefore, the first predetermined portion 51 and the third predetermined portion 53 of the liquid supply / drainage tube 50 are positioned on the same plane with respect to the container holding member 33.

[0041] A fourth predetermined portion 54 of the liquid supply / discharge tube 50, which is farther from the connection portion 50a than the second predetermined portion 52, is fixed by a fourth positioning member 64 so as not to move to the housing 10a of the cell culture device 10. That is, the liquid supply / discharge tube 50 is positioned at the first predetermined portion 51, the second predetermined portion 52, the third predetermined portion 53, and the fourth predetermined portion 54 which are spaced apart from one another in the longitudinal direction of the liquid supply / discharge tube 50.

[0042] The first positioning member 61 is rotatably attached to the front portion 33a of the container holding member 33. In Fig. 4, the first positioning member 61 rotates counterclockwise as a positive direction (+) rotation, and the first positioning member 61 rotates clockwise as a negative direction (-) rotation. Therefore, the first predetermined portion 51 of the liquid supply / drain tube 50 positioned by the first positioning member 61 can rotate along the surface of the container holding member 33 together with the first positioning member 61. That is, as shown in Fig. 4, the first predetermined portion 51 of the liquid supply / drain tube 50 can rotate 360 ​​degrees along the surface of the front portion 33a of the container holding member 33.

[0043] In contrast, the second positioning member 62 is fixed so as not to move relative to the front surface 32a of the turntable 32. Therefore, the second predetermined portion 52 of the liquid supply and drainage tube 50 positioned by the second positioning member 62 is immovable on the front surface 32a of the turntable 32. That is, as shown in Fig. 4, the second predetermined portion 52 of the liquid supply and drainage tube 50 is fixed to the turntable 32 in a state where it is disposed substantially parallel to the thickness direction of the front surface 32a of the turntable 32.

[0044] Similarly, the third positioning member 63 is fixed so as not to move relative to the mounting portion 33c of the container holding member 33. Therefore, the third predetermined portion 53 of the liquid supply and drainage tube 50 positioned by the third positioning member 63 is immovable on the surface of the mounting portion 33c of the container holding member 33. That is, as shown in Fig. 4, the third predetermined portion 53 of the liquid supply and drainage tube 50 is fixed to the container holding member 33 in a state where it is disposed substantially parallel to the mounting portion 33c of the container holding member 33 in the up-down direction.

[0045] In the cell culture device 10 of this embodiment, when supplying a reagent to the cell culture container 2 or draining a reagent from within the cell culture container 2, the turntable 32 and / or the container holding member 33 are rotated while the supply and drainage tube 50 is connected to the supply and drainage port 25, so that the supply and drainage tube 50 is slackened to have excess length between the first positioning member 61 attached to the container holding member 33 and the second positioning member 62 attached to the turntable 32, and between the second positioning member 62 attached to the turntable 32 and the fourth positioning member 64 attached to the surface of the housing of the cell culture device 10, A1.

[0046] That is, an excess portion N1 is formed between the first predetermined portion 51 and the second predetermined portion 52 of the liquid supply / drainage tube 50, and an excess portion N2 is formed between the second predetermined portion 52 and the fourth predetermined portion 54 of the liquid supply / drainage tube 50. As a result, even when the turntable 32 and / or the container holding member 33 are rotated, the liquid supply / drainage tube 50 is not pulled, and the rotation of the turntable 32 and / or the container holding member 33 is not restricted.

[0047] 7(a) shows a state in which the shelf surface 21b of the cell culture vessel 2 is arranged in a horizontal culture posture. When the vessel holding member 33 is rotated +90 degrees around the second axis Y by the second drive unit 35 from that state, the first predetermined portion 51 and the third predetermined portion 53 of the liquid supply and drainage tube 50 are positioned on the vessel holding member 33 by the first positioning member 61 and the third positioning member 63, respectively, as shown in FIG. 7(b), so that the length between the first predetermined portion 51 and the third predetermined portion 53 of the liquid supply and drainage tube 50 moves with the rotation of the vessel holding member 33 while being maintained substantially constant.

[0048] Further, the second predetermined portion 52 of the liquid supply / drainage tube 50 is positioned on the turntable 32 by the second positioning member 62, and the distance between the first positioning member 61 and the second positioning member 62 increases as the container holding member 33 rotates. Therefore, the slack of the extra length portion N1 formed between the first predetermined portion 51 and the second predetermined portion 52 of the liquid supply / drainage tube 50 changes to become less.

[0049] At this time, a force pulling the first predetermined portion 51 of the liquid supply / drainage tube 50, which is positioned in the container holding member 33 by the first positioning member 61, toward the second predetermined portion 52 acts on the first predetermined portion 51. Therefore, the first predetermined portion 51 of the liquid supply / drainage tube 50 rotates in the negative direction (-) along the surface of the container holding member 33.

[0050] 8(a) shows a state in which the shelf surface 21b of the cell culture vessel 2 is arranged in a horizontal culture posture. When the vessel holding member 33 is rotated −135 degrees around the second axis Y by the second drive unit 35 from that state, the first predetermined portion 51 and the third predetermined portion 53 of the liquid supply / discharge tube 50 are positioned on the vessel holding member 33 by the first positioning member 61 and the third positioning member 63, respectively, as shown in FIG. 8(b), so that the length between the first predetermined portion 51 and the third predetermined portion 53 of the liquid supply / discharge tube 50 moves with the rotation of the vessel holding member 33 while being maintained substantially constant.

[0051] Further, the second predetermined portion 52 of the liquid supply / drainage tube 50 is positioned on the turntable 32 by the second positioning member 62, and the distance between the first positioning member 61 and the second positioning member 62 increases as the container holding member 33 rotates. Therefore, the slack of the extra length portion N1 formed between the first predetermined portion 51 and the second predetermined portion 52 of the liquid supply / drainage tube 50 changes to become less.

[0052] At this time, a force pulling the first predetermined portion 51 of the liquid supply / drainage tube 50, which is positioned in the container holding member 33 by the first positioning member 61, toward the second predetermined portion 52 acts on the first predetermined portion 51. Therefore, the first predetermined portion 51 of the liquid supply / drainage tube 50 rotates in the positive direction (+) along the surface of the container holding member 33.

[0053] When rotating the cell culture vessel 2, it is necessary to rotate it at a speed that does not shake the culture medium and give excessive stimulation to the cells, and it is also necessary to supply the liquid evenly to each shelf 21. For this reason, the posture control is performed according to the following procedure.

[0054] (Fluid supply operation) From the state in which the shelf surface 21b of the cell culture vessel 2 is positioned in a horizontal culture position as shown in Figure 9(a), the controller C, which drives the drive units 34, 35, drives the first drive unit 34 to rotate the vessel holding member 33 and the cell culture vessel 2 by +90° around the first axis X so that the liquid supply / discharge port 25 is positioned downward as shown in Figure 9(b), and further drives the second drive unit 35 to rotate the cell culture vessel 2 by +90° around the second axis Y, thereby raising each shelf surface 21b and assuming a liquid supply position.

[0055] Then, after liquid is supplied to the horizontally oriented liquid supply / discharge ports 25 to allow the liquid to evenly enter each shelf space, the first drive unit 34 is driven to rotate the cell culture vessel 2 by -90° around the first axis X. At this time, when the liquid supply / discharge ports 25 become higher than the liquid level, the second drive unit 35 also begins to drive, rotating the cell culture vessel 2 by -90° around the second axis Y, bringing each shelf surface 21b into a horizontal culture position, and distributing the liquid approximately evenly over each shelf surface 21b.

[0056] (Drainage operation) In the drainage operation for draining the reagent, as shown in Fig. 10(a), from a state in which the shelf surface 21b of the cell culture vessel 2 is arranged in a horizontal culture posture, the turntable 32 and the vessel holding member 33 are first rotated +120° around the first axis X, and the vessel holding member 33 is rotated +30° around the second axis Y. Then, as shown in Fig. 10(b), the liquid supply / discharge port 25 faces downward, and the liquid in each shelf space flows out to the communication portion 21c and reaches the liquid supply / discharge port 25 (first liquid discharge posture). In this embodiment, the drainage operation for draining the reagent includes a drainage operation for draining a liquid such as a culture liquid.

[0057] 11(a) and 11(b) are diagrams for explaining the first drainage posture, and FIG. 11(b) is a diagram seen from the A direction of FIG. 11(a). In the first drainage posture of FIG. 11(a) and FIG. 11(b), if the diameter of the supply and drainage tube 50 is large, and the supply and drainage tube 50 has an upward gradient toward the downstream side, the liquid cannot rise all the way and remains in the region T1, which is the lowest point in FIG. 11(b). In that case, when a new reagent is supplied to the culture vessel, the used liquid is mixed in, which may affect the growth of cells. For example, in the process of supplying a detachment liquid, if there is any remaining liquid in the supply and drainage tube 50 when the medium of the previous process is drained, the detachment liquid is diluted, and detachment may not be performed properly.

[0058] One method for preventing liquid from remaining in the liquid supply / discharge tube 50 is to reduce the diameter of the liquid supply / discharge tube 50, but this would take a long time to deliver the liquid, and would lengthen the time the cells are suspended, which could affect the cells. If the flow rate is increased to shorten the liquid delivery time, excessive shear force would be applied to the cells, which could affect the cells.

[0059] Therefore, in the cell culture device of this embodiment, the drive mechanism 30 is appropriately controlled to rotate the turntable 32 and the container holding member 33 so that no liquid remains in the liquid supply / drainage tube 50 during drainage.

[0060] When the turntable 32 and the container holding member 33 are rotated by −220° around the first axis X and the container holding member 33 is rotated by −150° around the second axis Y from the first drainage posture in FIG. 11(a) and FIG. 11(b), the liquid supply / drain port 25 moves upward (second drainage posture). FIG. 12(a) to FIG. 12(c) are diagrams for explaining the second drainage posture, FIG. 12(b) is a diagram seen from the direction A in FIG. 12(a), and FIG. 12(c) is a diagram seen from the direction B in FIG. 12(a). Therefore, the region T1, which was the lowest point in FIG. 11(b), moves upward, and the liquid remaining in the region T1 moves toward the downstream side of the liquid supply / drain tube 50 (in the direction away from the connection portion 50a of the liquid supply / drain tube 50) to the region T2, which is the lowest point in FIG. 12(c).

[0061] When the turntable 32 and the container holding member 33 are rotated +190° around the first axis X and the container holding member 33 is rotated +120° around the second axis Y from the second drainage posture of Fig. 12(a) to Fig. 12(c), the liquid supply / drain port 25 moves downward (third drainage posture). Fig. 13(a) and Fig. 13(b) are diagrams for explaining the third drainage posture, and Fig. 13(b) is a view seen from the direction A in Fig. 13(a). Therefore, the region T2, which was the lowest point in Fig. 12(c), moves upward, and the liquid in the region T2 moves toward the region T3 on the downstream side of the liquid supply / drain tube 50, and all the liquid disappears from the liquid supply / drain tube 50.

[0062] In this way, when moving toward the downstream side of the liquid supply / drain tube 50, the liquid always flows toward the lowest point by not having the liquid supply / drain tube 50 slope upward toward the downstream side, thereby reducing the amount of liquid remaining in the liquid supply / drain tube 50. During the liquid drainage operation, the pump is constantly rotating to prevent the liquid from flowing back, and the liquid flows toward the downstream side of the liquid supply / drain tube 50. After drainage is completed, the liquid is returned to the liquid supply position of FIG. 9(b) for the next liquid supply.

[0063] As described above, the cell culture device 10 of this embodiment includes a base 31, a turntable 32 rotatably attached to the base 31 around a first axis X, a container holding member 33 rotatably attached to the turntable 32 around a second axis Y and holding a cell culture container 2, and a drive mechanism 30 that rotates the turntable 32 around the first axis X and rotates the container holding member 33 around the second axis Y. A supply and drainage port 25 to which a supply and drainage tube 50 is connected is arranged on the outer circumferential surface of the cell culture container 2. A first predetermined portion 51 of the supply and drainage tube 50, which is a predetermined length away from a connection portion 50a connected to the supply and drainage port 25, is positioned on the container holding member 33 by a first positioning member 61, and a second predetermined portion 52 of the supply and drainage tube 50, which is farther away from the connection portion 50a than the first predetermined portion 51, is positioned on the turntable 32 by a second positioning member 62. The first positioning member 61 is rotatably attached to the container holding member 33.

[0064] In this way, by rotatably attaching the first positioning member 61, which positions the first predetermined portion 51 of the liquid supply and drainage tube 50, to the container holding member 33, it is possible to eliminate the excess length formed between the first predetermined portion 51 and the second predetermined portion 52 of the liquid supply and drainage tube 50. Therefore, when supplying a reagent into the cell culture container 2 or discharging a reagent from within the cell culture container 2, even if the turntable 32 and / or the container holding member 33 are rotated while the liquid supply and drainage tube 50 is connected to the liquid supply and drainage port 25, it is possible to prevent the excess length of the liquid supply and drainage tube 50 connected to the liquid supply and drainage port 25 from becoming too long so that culture can be continued appropriately.

[0065] In the cell culture device 10 of this embodiment, the third predetermined portion 53 of the liquid supply / discharge tube 50 which is closer to the connection portion 50 a than the first predetermined portion 51 is positioned on the container holding member 33 by the third positioning member 63 .

[0066] In this way, when the turntable 32 and / or the container holding member 33 are rotated while the liquid supply and drainage tube 50 is connected to the liquid supply and drainage port 25, the direction of the first predetermined portion 51 of the liquid supply and drainage tube 50, which is rotatably positioned with respect to the container holding member 33 by the first positioning member 61, is easily changed. Therefore, excess length formed between the first predetermined portion 51 and the second predetermined portion 52 of the liquid supply and drainage tube 50 can be easily eliminated.

[0067] In the cell culture device 10 of this embodiment, the first predetermined portion 51 and the third predetermined portion 53 are positioned relative to the container holding member 33 on the same plane.

[0068] In this way, the direction of the first specified portion 51 of the liquid supply and drainage tube 50, which is rotatably positioned with respect to the container holding member 33 by the first positioning member 61, can be changed more easily. Therefore, the excess length formed between the first specified portion 51 and the second specified portion 52 of the liquid supply and drainage tube 50 can be further eliminated.

[0069] In the cell culture device 10 of this embodiment, during the drainage operation of draining reagent from the cell culture container 2, the drive mechanism 30 drives and rotates the turntable 32 and the container holding member 33 so that the reagent that has flowed out from the supply and drainage port 25 into the supply and drainage tube 50 flows through the supply and drainage tube 50 in a direction away from the supply and drainage port 25 due to gravity acting on it.

[0070] In this way, when the reagent is drained from the cell culture vessel 2, the reagent can be prevented from remaining in the supply and drainage tube 50.

[0071] Although the embodiment of the present invention has been described above, the specific configuration of each part is not limited to the above-described embodiment.

[0072] For example, in the above embodiment, the first predetermined portion 51 is positioned to the right of the opening 33b near the upper end of the front portion 33a of the container holding member 33, and the second predetermined portion 52 is positioned to the front portion 32a of a plate-like member that faces the right side portion of the container holding member 33 on the turntable 32, but is not limited to this. The location where the first predetermined portion 51 is positioned on the container holding member 33 and the location where the second predetermined portion 52 is positioned on the turntable 32 are arbitrary.

[0073] In the above embodiment, the third predetermined portion 53, which is closer to the connection portion 50a than the first predetermined portion 51 of the liquid supply / drainage tube 50, is positioned in the container holding member 33 by the third positioning member 63, but this is not limited to the above. The portion between the connection portion 50a and the first predetermined portion 51 of the liquid supply / drainage tube 50 does not have to be positioned in the container holding member 33. Furthermore, when the portion between the connection portion 50a and the first predetermined portion 51 of the liquid supply / drainage tube 50 is positioned in the container holding member 33, the number of positions to be positioned is arbitrary.

[0074] In the above embodiment, the first specified portion 51 of the liquid supply / drainage tube 50 is attached to the container holding member 33 so as to be rotatable 360 ​​degrees, but this is not limited thereto. The range within which the first specified portion 51 can rotate with respect to the container holding member 33 is arbitrary.

[0075] In the above embodiment, the second predetermined portion 52 of the liquid supply / drain tube 50 is fixed so as not to move relative to the turntable 32, but the second predetermined portion 52 may be attached rotatably relative to the turntable 32. Similarly, in the above embodiment, the third predetermined portion 53 of the liquid supply / drain tube 50 is fixed so as not to move relative to the container holding member 33, but the third predetermined portion 53 may be attached rotatably relative to the container holding member 33. In the above embodiment, not only the first positioning member 61 but also any of the first positioning member 61, the second positioning member 62, and the third positioning member 63 may be rotatable relative to the turntable 32 or the container holding member 33.

[0076] In the above embodiment, the first predetermined portion 51 and the third predetermined portion 53 are positioned on the same plane with respect to the container holding member 33, but this is not limited thereto. The first predetermined portion 51 and the third predetermined portion 53 may be positioned on different planes.

[0077] In the above embodiment, the liquid supply / drain port 25 and the liquid supply / drain tube 50 are used for both liquid supply and liquid drainage, but this is not limited to the above. Different liquid ports and liquid tubes may be used for liquid supply and liquid drainage, respectively, to separate the liquid supply path and the liquid drainage path.

[0078] In the above embodiment, in the drainage operation for draining the reagent from the cell culture vessel 2, the turntable 32 and the container holding member 33 are rotationally driven so as to change to the first drainage posture, the second drainage posture, and the third drainage posture in this order, but this is not limited thereto. In the drainage operation, as long as the turntable 32 and the container holding member 33 are rotationally driven so that the reagent that has flowed out from the supply / drain port 25 into the supply / drain tube 50 flows in the supply / drain tube 50 in a direction away from the supply / drain port 25 due to gravity acting thereon, the procedure for changing the posture of the turntable 32 and the container holding member 33 may be a procedure other than the above.

[0079] Furthermore, even if the first specified portion 51 of the supply and drainage tube 50 is fixed so as not to rotate relative to the container holding member 33, during the drainage operation, it is possible for the turntable 32 and the container holding member 33 to be rotated so that the reagent that has flowed out from the supply and drainage port 25 into the supply and drainage tube 50 flows within the supply and drainage tube 50 in a direction away from the supply and drainage port 25 due to gravity acting on it.

[0080] In the above embodiment, a cell culture vessel 2 having a multi-tiered shelf structure is used, but a cell culture vessel 2 not having a multi-tiered shelf structure may be used. In the above embodiment, the liquid supply / discharge tube 50 is not limited to one through which a liquid such as a reagent or culture solution flows, and may be one through which a gas (air) or the like flows.

[0081] Other configurations can also be modified in various ways without departing from the spirit of the present invention. [Explanation of symbols]

[0082] 2 Cell culture vessel 10 Cell culture equipment 25 Liquid supply / drain port (liquid port) 30 Drive mechanism 31 Base 32 Rotating table 33 Container holding member 50 Liquid supply / drainage tube (liquid tube) 50a Connection 51 First Specified Section 52 Second Specified Section 53 Third Specified Section 61 First positioning member 62 Second positioning member 63 Third positioning member X…1st axis Y...Second axis

Claims

1. With the base, a rotating table rotatably attached to the base around a first axis; a container holding member that is rotatably attached to the turntable around a second axis and holds a cell culture container; a drive mechanism that rotates the rotary table about a first axis and rotates the container holding member about a second axis, A liquid port to which a liquid tube is connected is arranged on an outer peripheral surface of the cell culture vessel, a first predetermined portion of the liquid tube, the first predetermined portion being spaced a predetermined length from a connection portion connected to the liquid port, is positioned on the container holding member by a first positioning member; a second predetermined portion of the liquid tube that is farther from the connection portion than the first predetermined portion is positioned on the rotating table by a second positioning member; the first positioning member is rotatably attached to the container holding member, A cell culture device characterized in that when the first positioning member that positions the first specified portion rotates relative to the container holding member, the orientation of the liquid tube upstream of the first specified portion and the orientation of the liquid tube downstream of the first specified portion change.

2. With the base, a rotating table rotatably attached to the base around a first axis; a container holding member that is rotatably attached to the turntable around a second axis and holds a cell culture container; a drive mechanism that rotates the rotary table about a first axis and rotates the container holding member about a second axis, A liquid port to which a liquid tube is connected is arranged on an outer peripheral surface of the cell culture vessel, a first predetermined portion of the liquid tube, the first predetermined portion being spaced a predetermined length from a connection portion connected to the liquid port, is positioned on the container holding member by a first positioning member; a second predetermined portion of the liquid tube that is farther from the connection portion than the first predetermined portion is positioned on the rotating table by a second positioning member; The cell culture device, characterized in that the first positioning member positions the first specific portion on the container holding member at a position not on the second axis and is rotatably attached to the container holding member.

3. With the base, a rotating table rotatably attached to the base around a first axis; a container holding member that is rotatably attached to the turntable around a second axis and holds a cell culture container; a drive mechanism that rotates the rotary table about a first axis and rotates the container holding member about a second axis, A liquid port to which a liquid tube is connected is arranged on an outer peripheral surface of the cell culture vessel, a first predetermined portion of the liquid tube, the first predetermined portion being spaced a predetermined length from a connection portion connected to the liquid port, is positioned on the container holding member by a first positioning member; a second predetermined portion of the liquid tube that is farther from the connection portion than the first predetermined portion is positioned on the rotating table by a second positioning member; a third predetermined portion of the liquid tube, the third predetermined portion being closer to the connection portion than the first predetermined portion, is positioned on the container holding member by a third positioning member; the first positioning member is rotatably attached to the container holding member, The cell culture device according to claim 1, wherein the first and third predetermined portions are positioned on the same plane relative to the container holding member.

4. The cell culture device described in any one of claims 1 to 3, characterized in that, during a drainage operation to drain reagent from the cell culture container, the driving mechanism rotates the turntable and the container holding member so that the reagent flowing out from the liquid port into the liquid tube flows through the liquid tube in a direction away from the liquid port due to gravity acting on it.

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