Cell culture system and cell culture device

Abstract

To provide a cell culture system that does not require a high-grade clean environment and can appropriately handle multi-shelf type cell culture vessels even under many space constraints.SOLUTION: Provided is a cell culture system CCS configured so that a space between a cell culture vessel 2, a refrigerator 1 for storing a liquid such as a medium and a reagent to be supplied to the cell culture vessel 2, and a cell recovery section 3 for recovering cells cultured in the cell culture vessel 2 becomes a spatially closed line L spatially closed at least from when the liquid is transferred to the cell culture vessel 2 until when the cultured cells are removed. The cell culture vessel 2 is arranged in an incubator 4 as a multi-shelf type having a liquid supply and drainage port. This allows the cell culture system CCS to change in posture within the incubator 4 between a posture where the liquid is distributed to each shelf through the liquid supply and drainage port while maintaining a closed line state, and a posture where the liquid is collected and drained from each shelf through the liquid supply and drainage port.SELECTED DRAWING: Figure 1

Description

[Technical field]

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

[0002] In recent years, a cell culture technique has become known in which cells seeded in a culture solution are cultured using a multi-layered cell culture vessel containing multiple trays in order to culture a large amount of cells. In this type of cell culture technique, in order to reduce the burden on the operator, a cell culture device is used that performs handling operations such as holding and rotating the cell culture vessel in order to introduce the culture solution in which the cells are seeded into the cell culture vessel or to recover the culture solution from the cell culture vessel (see Patent Document 1).

[0003] This is a cell culture system having a cart device that can move while carrying a multi-layered cell culture vessel containing multiple trays, and an operating device that can hold and rotate the cell culture vessel, wherein the cart device has a cart with wheels and a fixing member that is mounted on the cart so as to be freely attached and detached to the cart and fixes the cell culture vessel to the cart, and the operating device is configured with a rotating unit that holds the cell culture vessel together with the fixing member and performs a rotational operation to rotate the cell culture vessel about a first rotation axis and / or a second rotation axis.

[0004] Once cell culture is complete, the cell culture vessel is removed from the device and transported on the cart to another location to collect the cells. Next, a suspension of cells and culture medium that needs to be cultured is placed into the cell culture vessel, which is then transported again on the cart and set in the device. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2018-138056 A Summary of the Invention [Problem to be solved by the invention]

[0006] However, in this configuration, there is a problem that the temperature of the cells is lowered when the cell culture vessel is taken out of the incubator, stimulating the cells. In addition, when the cell culture vessel is taken out of the incubator and transported, the culture medium is shaken, which may impact the cells and cause them to peel off or be stimulated. Furthermore, when using a multi-layer culture vessel, the vessel is heavy, and it is not easy to transport it manually. Furthermore, there is a problem that the cells must be discarded due to contamination, etc., when the tubes for delivering the liquid are reconnected.

[0007] Therefore, in order to replace the culture medium inside the incubator, it is conceivable to configure the incubator by bringing a scaled-down version of the configuration of Patent Document 1 into the incubator. However, even inside the incubator, changing the connections of tubes can cause contamination due to the introduction of bacteria. Therefore, it is necessary to at least maintain a clean atmosphere inside the incubator, and considering the opening and closing of the incubator door, a high level of cleanliness is ultimately required for the room in which the incubator is installed.

[0008] When attaching or detaching tubes to cell culture vessels in an incubator, CO2 gas and other gases are introduced inside, causing the gas to leak and humidify the room, which can lead to rust. For this reason, humidity control is also required.

[0009] Furthermore, in order to handle multi-layer cell culture vessels in an incubator, which is a space with many restrictions, appropriate ingenuity is required.

[0010] The present invention focuses on these problems and aims to realize a novel cell culture system and cell culture device that does not require a high-grade clean environment and can properly handle multi-layered cell culture vessels even with many space constraints. [Means for solving the problem]

[0011] In order to achieve the above object, the present invention takes the following measures.

[0012] That is, the cell culture system of the present invention is configured so that a spatially closed line is formed between a cell culture vessel, a liquid storage section for storing liquid such as culture medium and reagents to be supplied to the cell culture vessel, and a cell recovery section for recovering cells cultured in the cell culture vessel, at least from the time when liquid is transferred to the cell culture vessel to the time when the cultured cells are removed, and is characterized in that the cell culture vessel is arranged in an incubator in a multi-tiered shelf type having a multi-layered structure with supply and drainage ports, and the position of the cell culture vessel can be changed within the incubator between a position in which liquid is evenly spread and distributed to each layer of shelves through the supply and drainage ports while maintaining the closed line state, and a position in which liquid is collected and drained from each shelf through the supply and drainage ports.

[0013] In this way, the use of a multi-shelf cell culture vessel makes it possible to handle large-volume culture. In this case, since the cell culture vessel is not taken out of the incubator, the cells are not damaged by temperature changes or vibrations, and since no manual procedures are required, the labor is reduced. Furthermore, since a closed system is maintained, contamination due to the introduction of bacteria can be prevented, and since it is not necessary to maintain a high level of cleanliness in the incubator and the room, the labor of wearing excessive clean clothes when operating the device can be eliminated. Similarly, since gas does not leak into the incubator because of the closed system line, there is no need to control the humidity inside the incubator. Furthermore, since the number of supply and discharge ports can be reduced by distributing and collecting liquid from the supply and discharge ports to each shelf, the peripheral structure of the cell culture vessel that moves within the incubator can be simplified.

[0014] In this case, by placing a plurality of the cell culture vessels in an incubator and connecting at least the cell culture vessels to each other via a closed line so as to enable subculturing of the cultured cells, it is possible to significantly increase the culture rate while maintaining the basic effect of the present invention.

[0015] The cell culture device of the present invention is characterized in that it comprises a multi-shelf cell culture vessel arranged in an incubator and having supply and drainage ports, a drive mechanism that enables the cell culture vessel to be changed between a position in which liquid is distributed to each shelf through the supply and drainage ports and a position in which liquid on each shelf is collected and drained through the supply and drainage ports, and a supply and drainage tube that remains connected to the supply and drainage ports during the drive.

[0016] In this way, the number of movable supply and discharge tubes connected to the cell culture vessel can be minimized, while the cell culture vessel can be appropriately handled.

[0017] In this case, the supply and drainage ports can be composed of a supply port and a drainage port, to which a supply tube and a drainage tube can be connected, respectively. However, if the supply and drainage ports serve both as supply ports for supplying liquid to each shelf and as drainage ports for discharging liquid from each shelf, and the driving mechanism drives the cell culture vessel so that it assumes a different posture when supplying and draining liquid, the number of ports and tubes can be further reduced.

[0018] In the above, the cell culture vessel has a communication portion to each shelf space on one edge side of each shelf forming a multi-tiered shelf structure, each shelf space is separated from the other portions except for the communication portion, and the one edge is provided with a vertical wall that can store liquid on the shelf surface when the shelf is horizontal, and the drive mechanism is effective to be configured to drive the cell culture vessel to sequentially assume a liquid supply posture in which each shelf surface is raised so that the communication portion is located at the bottom and liquid enters each shelf evenly, an intermediate posture in which the cell culture vessel is rotated from that state so that the communication portion is located at the top and liquid in each shelf space is separated, and a culture posture in which the shelf surface of each shelf is subsequently rotated so that it is horizontal and liquid is distributed approximately evenly over each shelf surface.

[0019] If the cell culture vessel is immediately shifted from the liquid supplying position to the culture position, the lower shelves will be more likely to receive liquid and the upper shelves will be less likely to receive liquid, resulting in an uneven state. To prevent this, the cell culture vessel must be rotated quickly. This can cause damage to the cells and can lead to drying of the cells. On the other hand, if the intermediate position is inserted as described above, the shelves will not be vertically separated from the liquid supplying position to the intermediate position without the need to rotate the cell culture vessel quickly, and the liquid will be evenly separated into each shelf space. Therefore, the liquid that has entered each shelf space can be stored on the horizontal shelf surface without the need to rotate quickly from the intermediate position to the culture position. As a result, the cell culture vessel can be changed in position at an appropriate speed, reducing damage to the cells and preventing them from drying out.

[0020] Furthermore, the drive mechanism effectively includes a first drive unit that rotates the cell culture vessel around a first axis, a second drive unit that rotates the cell culture vessel around a second axis perpendicular to the first axis, and a control unit that controls these drive units, wherein the first drive unit is positioned in a direction along the first axis, and the second drive unit is disposed near the first drive unit and is connected to the second axis via a transmission unit that transmits power along a direction parallel to the first axis.

[0021] In this way, at least the drive mechanism does not become bulky in the second axial direction, so that it can be appropriately arranged inside an incubator which has significant spatial limitations.

[0022] Furthermore, if a cell culture vessel that changes its posture with the supply and drainage tube connected is further equipped with an automatic winding mechanism that winds up the excess length of the supply and drainage tube while applying a certain tension to the tube, the supply and drainage tube can be effectively prevented from becoming tangled with its surroundings even when the cell culture vessel is rotated with the supply and drainage tube connected. Effect of the Invention

[0023] Because of the configuration described above, the present invention makes it possible to provide a new and useful cell culture system and cell culture device that does not require a high-grade clean environment and can appropriately handle multi-shelf-type cell culture vessels even under many space constraints. [Brief description of the drawings]

[0024] [Figure 1] FIG. 1 is a schematic configuration diagram of a cell culture system according to one embodiment of the present invention. [Diagram 2] FIG. 2 is a diagram showing an incubator housing a cell culture vessel in the system. [Diagram 3] FIG. 2 is a diagram showing the shelf structure of the cell culture vessel. [Figure 4] FIG. 2 is a diagram showing a cell culture vessel together with a driving mechanism. [Diagram 5] FIG. 2 is a diagram showing a cell culture vessel together with a driving mechanism. [Figure 6] FIG. 2 is a diagram showing a cell culture vessel together with a driving mechanism. [Figure 7] 4A to 4C are diagrams showing a driving procedure of the cell culture vessel. [Figure 8] 4A to 4C are diagrams showing a driving procedure of the cell culture vessel. [Figure 9] 4A to 4C are diagrams showing a driving procedure of the cell culture vessel. [Figure 10] 4A to 4C are diagrams showing a driving procedure of the cell culture vessel. [Figure 11] 4A to 4C are diagrams showing a driving procedure of the cell culture vessel. [Figure 12] FIG. [Figure 13] FIG. 13 is a diagram showing another modified example of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

[0026] 1 is a system diagram showing a simplified view of the cell culture system CCV of this embodiment. This cell culture system CCV includes a cell culture vessel 2, a refrigerator 1 serving as a liquid storage section for storing liquids such as culture medium and various reagents to be supplied to the cell culture vessel 2, and a cell recovery section 3 for recovering cells cultured in the cell culture vessel 2. The cell culture vessel 2, the refrigerator 1, 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 L until operation is terminated, preventing the intrusion of bacteria from the outside.

[0027] The inside of refrigerator 1 is kept at a temperature required for keeping culture media and various reagents cool, for example, at about 5°C.

[0028] 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.

[0029] 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 IP 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 IP are also composed of the line L of a closed system.

[0030] In such a closed line L, a system is constructed which performs the following processes all automatically without human intervention: preparing liquids such as culture medium and reagents in refrigerator 1, transferring those liquids from refrigerator 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 on the liquid removed from cell culture vessel 2 before recovery in intermediate processing section IP.

[0031] For this purpose, valves V and pumps P (pressure pump P(a) and suction pump P(b)) are connected to each line L as necessary, and these valves V and pumps P are controlled by control commands SG from a controller CT.

[0032] In this embodiment, in order to maintain the cell culture vessel 2 at a predetermined temperature without breaking the closed system line L, the cell culture device CV including the cell culture vessel 2 and peripheral components is placed in the incubator 4 as shown in Figures 1 and 2 with the supply and discharge tube 61 that forms part of the closed system line L still connected.

[0033] 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.

[0034] 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 supply from the outside is distributed to 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 L as much as possible, a liquid supply and drainage port 22 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 22 is common to each shelf 21, and the cell culture vessel 2 is changed in position in the incubator 4 while maintaining a closed line state, thereby realizing the supply of liquid to each shelf 21 and the drainage from each shelf 21.

[0035] 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 a corner part C on one edge 21a side of each shelf 21 so that liquid can flow evenly into each shelf space S, and the shelf spaces S 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.

[0036] The cell culture device CV includes a drive mechanism 5 for changing the attitude of the cell culture vessel 2, as shown in FIGS.

[0037] The drive mechanism 5 comprises a base 51, a rotating table 52 rotatably attached to the base 51 about a first axis X, a container holding member 53 rotatably attached to the rotating table 52 about a second axis Y, a first drive unit 54 that drives and rotates the rotating table 52 about the first axis X, a second drive unit 55 that drives and rotates the container holding member 53 about the second axis Y, and the controller CT which serves as a control unit for controlling these drive units 54, 55. The cell culture container 2 can be detachably attached to the container holding member 53 via a clamping unit 53a that utilizes elastic force.

[0038] The first drive unit 54 is located in a direction along the first axis X from the cell culture vessel 2, and the second drive unit 55 is disposed near the first drive unit 54 and is connected to the second axis Y via a transmission mechanism 56 (see FIG. 7) using a belt, chain, or the like that transmits power along the first axis X.

[0039] That is, when only the first driving unit 54 is driven, the turntable 52, the container holding member 53, and the cell culture vessel 2 rotate around the first axis X as shown in Fig. 7, and when only the second driving unit 55 is driven, the turntable 52 does not rotate but the container holding member 53 and the cell culture vessel 2 rotate around the second axis Y as shown in Fig. 6, and further, when the first driving unit 54 and the second driving unit 55 are driven simultaneously, the turntable 52, the container holding member 53, and the cell culture vessel 2 rotate around the first axis X as shown in Fig. 7, and further, the container holding member 53 and the vessel 2 can rotate around the second axis Y relative to the turntable 52 as shown in Fig. 6. The driving mechanism 5 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.

[0040] 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.

[0041] First, the controller CT in Fig. 1, which drives the drive units 54 and 55, drives the first drive unit 54 to rotate the cell culture vessel 2 by 90° around the first axis X so that the communication unit 21c is positioned downward as shown in Fig. 7, and each shelf surface 21b is raised to a liquid supplying position. Then, liquid is supplied to the liquid supply / discharge port 22 facing horizontally from the front to the back of the page in Fig. 7, so that the liquid evenly enters each shelf space S. At this stage, liquid can move between each shelf space S when the position of the cell culture vessel 2 changes.

[0042] From that state, the second drive unit 55 is driven to rotate the cell culture vessel 2 90° around the second axis Y (see FIG. 8), and the first drive unit 54 is further driven to rotate the cell culture vessel 2 90° around the first axis X (see FIG. 9) to bring the cell culture vessel 2 into the intermediate position. As a result, the communication portion 21c is positioned at the top, and the liquid is divided approximately equally into each shelf space S. At this stage, the liquid that has entered each shelf space S cannot move back and forth even if the position of the cell culture vessel 2 changes.

[0043] Furthermore, the second drive unit 55 is driven to rotate the cell culture vessel 2 by 90° around the second axis Y, so that each shelf surface 21b is placed in a horizontal culture posture, and the liquid is spread approximately evenly over each shelf surface 21b (see FIG. 10). The liquid is stored in the hatched area in FIG. 10(b), and the culture is carried out in this state.

[0044] 11, when draining liquid, the first drive unit 54 is driven about the first axis X to an angle θ1 with respect to the horizontal, and the second drive unit 55 is driven about the second axis Y to an angle θ2 with respect to the horizontal so that the liquid supply / drain port 22 faces downward and the liquid in each shelf space S comes out of the communication portion 21c and reaches the liquid supply / drain port 22 (liquid draining posture). θ1 and θ2 are set to, for example, about 30° so that no liquid remains.

[0045] After draining, return to the position shown in Figure 7 for the next supply of fluid.

[0046] The controller CT shown in FIG. 1 incorporates sequences related to the driving by the series of driving units 54, 55, the pumps P (P(a), P(b)) for supplying and discharging liquid, and the valves V at each portion.

[0047] During the culture, the medium needs to be replaced at a predetermined frequency (for example, once a day). As shown in Figs. 1 and 2, a plurality of cell culture vessels 2(A) and 2(B) (two in the illustrated example) may be installed in an incubator 4, and a drive mechanism 5 and a liquid supply and drainage tube 61 may be connected to each of them, and the cells cultured in the first vessel may be transferred to the second vessel for further proliferation, that is, a so-called subculture may be performed. For this reason, the controller CT shown in Fig. 1 also incorporates a sequence for replacing the medium for each of the cell culture vessels 2(A) and 2(B), a sequence for detaching the cells cultured in the first cell culture vessel 2(A) and draining them from the liquid supply and drainage port 22, and introducing them from the liquid supply and drainage port 22 of the second cell culture vessel 2(B), and the like.

[0048] In each drawing, reference numeral 23 denotes a supply and exhaust port, which allows gas to be released from inside when supplying liquid through the supply and exhaust port 22, and allows gas to be introduced inside when discharging liquid through the supply and exhaust port 22, thereby ensuring smooth supply and drainage. Reference numeral 62 denotes a supply and exhaust tube. The supply and exhaust port 22 is inserted and removed from the bottom of the cell culture vessel 2, thereby preventing gas bubbles from being mixed into the liquid.

[0049] In the series of operations, the drive of the cell culture vessel 2 by the drive units 54, 55 is controlled so as not to rotate the cell culture vessel 2 by more than 180° around the X-axis and Y-axis, so as to prevent the supply and discharge tube 61 and the supply and exhaust tube 62 from becoming entangled. In the figure, reference numeral 21c' denotes a communication portion that is provided by cutting out each shelf surface 21b and communicates with the supply and exhaust port 23, and reference numeral 21e' denotes a standing wall for storing liquid on each shelf surface 21b.

[0050] As described above, the cell culture system CCS of this embodiment is configured so that a spatially closed line L is formed between the cell culture vessel 2, the refrigerator 1 which serves as a liquid storage section for storing liquids such as culture medium and reagents to be supplied to the cell culture vessel 2, and the cell recovery section 3 which recovers the cells cultured in the cell culture vessel 2, at least from the time when the liquid is transferred to the cell culture vessel 2 to the time when the cultured cells are removed.The cell culture vessel 2 is arranged in an incubator 4 in a multi-shelf type having a supply and drainage port 22, and is characterized in that the position of the cell culture system CCS can be changed within the incubator 4 between a position in which liquid is distributed to each shelf 21 through the supply and drainage port 22 while maintaining a closed line state, and a position in which liquid is collected and drained from each shelf 21 through the supply and drainage port 22.

[0051] In this way, by using the multi-shelf type cell culture vessel 2, it is possible to handle large-volume culture. In this case, since the cell culture vessel 2 is not taken out of the incubator 4, the cells are not damaged by temperature changes or vibrations, and since no manual procedures are required, the labor is reduced. Furthermore, since a closed system is maintained, contamination due to the inclusion of bacteria can be prevented, and since it is not necessary to maintain a high level of cleanliness inside the incubator 4 and the room, the labor of wearing excessive clean clothes when operating the device can be eliminated. Similarly, since gas does not leak into the incubator 4 because of the closed system line L, there is no need to control the humidity inside the incubator 4. Furthermore, since the number of supply and discharge ports 22 can be reduced by distributing and collecting liquid from the supply and discharge ports 22 to each shelf 21, the peripheral structure of the cell culture vessel 2 movable inside the incubator 4 can be simplified.

[0052] Furthermore, multiple cell culture vessels 2 (2(A), 2(B)) are placed in the incubator 4, and at least the cell culture vessels 2(A), 2(B) are connected to each other via a closed line L so that the cultured cells can be passaged. This makes it possible to significantly increase the culture rate while maintaining the basic effects of this embodiment.

[0053] On the other hand, the cell culture device CV of this embodiment is equipped with a multi-shelf cell culture vessel 2 arranged in an incubator 4 and having supply and drainage ports 22, a drive mechanism 5 that can change the position of this cell culture vessel 2 between a position in which liquid is distributed to each shelf 21 through the supply and drainage ports 22 and a position in which liquid is collected from each shelf 21 and drained through the supply and drainage ports 22, and a supply and drainage tube 61 that remains connected to the supply and drainage port 22 during the drive, so that the number of supply and drainage tubes 61 that are movable while connected to the cell culture vessel 2 can be minimized while appropriate handling of the cell culture vessel 2 can be performed.

[0054] In a configuration not shown, the supply and drainage port 22 can be configured as a supply port and a drainage port, to which a supply tube and a drainage tube can be connected, respectively; however, in this embodiment, the supply and drainage port 22 serves both as a supply port for supplying liquid to each shelf 21 and as a drainage port for discharging liquid from each shelf 21, and the driving mechanism 5 is configured to drive the cell culture vessel 2 so that its posture is different when supplying and draining liquid, thereby further reducing the number of ports and tubes.

[0055] In this case, the cell culture vessel 2 has a communication part 21c with each shelf space S on one end edge 21a side of each shelf forming a multi-tiered shelf structure, and the shelf spaces S are partitioned apart from each other except for the communication part 21c, and the one end edge 21a is provided with a standing wall 21e that can store liquid on the shelf surface 21b when the shelf 21 is horizontal. The drive mechanism 5 is configured to drive the cell culture vessel 2 to sequentially assume a liquid supply posture (see FIG. 7) in which each shelf 21 is raised so that the communication part 21c is located at the bottom and liquid is evenly introduced into each shelf surface 21b, an intermediate posture (see FIG. 9) in which the cell culture vessel 2 is rotated from that state so that the communication part 21c is located at the top and liquid in each shelf space S is separated, and a culture posture (see FIG. 10) in which the shelf surface 21b of each shelf 21 is rotated so that it is horizontal and liquid is spread almost evenly on each shelf surface 21b.

[0056] That is, when the cell culture vessel 2 is immediately rotated around the first axis X from the liquid supplying posture shown in FIG. 7 to the culture posture shown in FIG. 10, an uneven state occurs in which the lower shelves 21 are more likely to receive liquid and the upper shelves 21 are less likely to receive liquid over time, so that the cell culture vessel 2 must be rotated quickly to prevent this. This may cause damage to the cells. On the other hand, when the intermediate posture as described above is interposed, the shelves 21 are not vertically separated from each other from the liquid supplying posture shown in FIG. 7 to the intermediate posture shown in FIG. 9 without the need to rotate the cell culture vessel 2 quickly, and the liquid is uniformly separated into each shelf space S. Therefore, the liquid that has entered each shelf space S can be transferred to a state in which it is stored on the horizontal shelf surface 21b in the same amount without the need to rotate quickly from the intermediate posture to the culture posture. As a result, the posture of the cell culture vessel 2 can be changed at an appropriate speed, and damage to the cells can be reduced.

[0057] The driving mechanism 5 includes a first driving unit 54 that drives and rotates the cell culture vessel 2 around a first axis X, a second driving unit 55 that drives and rotates the cell culture vessel 2 around a second axis Y perpendicular to the first axis X, and a controller CT that serves as a control unit for controlling these driving units 54 and 55. The first driving unit 54 is located in a direction along the first axis X, and the second driving unit 55 is disposed near the first driving unit 54 and is connected to the second axis Y via a transmission unit 56 that transmits power along a direction parallel to the first axis X. Since the driving mechanism 5 is not bulky at least in the direction of the second axis Y, it can be appropriately disposed in the incubator 4, which has significant spatial constraints.

[0058] Although one embodiment of the present invention has been described above, the specific configuration of each part is not limited to the above-described embodiment.

[0059] For example, for a cell culture vessel 2 that changes position while the supply and drainage tube 61 is connected, it may be effective to further provide an automatic winding mechanism 7 that winds up the excess length of the supply and drainage tube 61 while applying a certain tension to the tube, as shown in FIG. 12.

[0060] This prevents the liquid supply and drainage tube 61 from drooping and becoming tangled with the surroundings. In addition, since the liquid supply and drainage tube 61 does not droop, it is possible to prevent liquid from remaining in the drooping portion of the liquid supply and drainage tube 61. As the automatic winding mechanism 7, for example, there is one in which the liquid supply and drainage tube 61 is wound around a rotating mechanism equipped with an actuator, and the actuator of the rotating mechanism is rotated forward and backward in accordance with the angle at which the cell culture vessel 2 is turned to extend and retract the liquid supply and drainage tube, thereby maintaining a constant tension regardless of the angle at which the cell culture vessel 2 is turned.

[0061] Furthermore, there is no prohibition on configuring the liquid supply / drainage ports and the liquid supply / drainage tubes separately for liquid supply and liquid drainage.

[0062] Furthermore, the configuration of the drive mechanism and the procedure for changing the posture of the cell culture vessel may be configured and performed in a manner other than those described above, as long as the intermediate posture can be reached.

[0063] Furthermore, in the above embodiment, the refrigerator 1 serving as a liquid storage unit is disposed externally, and the liquid in the refrigerator 1 is directly supplied to the cell culture vessels 2 (2(A), 2(B)) in the incubator 4. However, as shown in FIG. 13, a bag 101 serving as a liquid storage unit may be disposed in the incubator 4, for example by hanging it, and liquid may be supplied from the bag 101 to the cell culture vessels 2 (2(A), 2(B)).

[0064] In this case, as shown in FIG. 13(a), the liquid from the refrigerator 1 can be temporarily stored in a bag 101 and then supplied to the cell culture container 2 (2(A), 2(B)), or as shown in FIG. 13(b), there can be no external refrigerator 1 and only liquid is supplied from the bag 101.

[0065] 13(a), it is also effective to provide a preheating unit 101a that preheats the liquid before it is introduced into the incubator 4. If a low-temperature solution is placed in the cell culture vessel 2 (2(A), 2(B)) without preheating, there is a risk that it may stimulate the cells. However, if the solution is preheated in advance, it is possible to prevent the temperature inside the incubator 4 from fluctuating, and it becomes possible to appropriately control the temperature inside the incubator 4.

[0066] Such a preheating unit 101a can of course be effectively applied to the configuration of FIG. 1, and by preheating the liquid before it is introduced into the incubator 4, it becomes possible to optimize the temperature control.

[0067] Of course, a configuration in which the liquid introduced into the incubator 4 is heated in the incubator 4 and supplied to the cell culture vessels 2 (2(A), 2(B)) can also be appropriately adopted.

[0068] Other configurations can also be modified in various ways without departing from the spirit of the present invention. [Explanation of symbols]

[0069] 1...Liquid storage section (refrigerator) 2, 2(A), 2(B)...Cell culture container 3. Cell collection section 4. Incubator 5...Drive mechanism 7…Automatic winding mechanism 21…Shelf 21a...One edge 21b…Shelf surface 21c...Communication part 21e…standing wall 22...Fluid supply / drain port 54...First drive mechanism 55...Second drive mechanism 101...Liquid storage section (bag) CCS…Cell Culture System CD…Cell culture device CT: Control unit (controller) L: Closed system line S…Shelf space X…1st axis Y...Second axis

Claims

1. a cell culture vessel, a liquid storage section for storing a liquid such as a medium or a reagent to be supplied to the cell culture vessel, and a cell recovery section for recovering cells cultured in the cell culture vessel are configured to form a spatially closed line system at least during the period from the transfer of the liquid to the cell culture vessel to the removal of the cultured cells; The cell culture system is characterized in that the cell culture vessels are arranged in a multi-shelf type having supply and drainage ports within an incubator, and the position within the incubator can be changed between a position in which liquid is distributed to each shelf through the supply and drainage ports while maintaining the closed line state, and a position in which liquid is collected and drained from each shelf through the supply and drainage ports.

2. 2. The cell culture system according to claim 1, wherein a plurality of the cell culture vessels are arranged in an incubator, and at least the cell culture vessels are connected to each other via a closed line so as to enable subculture of the cultured cells.

3. A cell culture device comprising: a multi-shelf cell culture vessel arranged in an incubator and having supply and drainage ports; a drive mechanism that can change the position of the cell culture vessel between a position in which liquid is distributed to each shelf through the supply and drainage ports and a position in which liquid on each shelf is collected and drained through the supply and drainage ports; and a supply and drainage tube that remains connected to the supply and drainage ports during the drive.

4. The cell culture device according to claim 3, wherein the supply and drainage ports serve as supply ports for supplying liquid to each shelf and as drainage ports for discharging liquid from each shelf, and the drive mechanism drives the cell culture vessel so that the posture is different when supplying and draining liquid.

5. 5. The cell culture device according to claim 4, wherein the cell culture vessel has a communication portion with each shelf space on one edge side of each shelf forming a multi-tiered shelf structure, each shelf space is separated from the other in the portions other than the communication portion, and the one edge is provided with a standing wall capable of storing liquid on the shelf surface when the shelf is horizontal, and the drive mechanism drives the cell culture vessel to sequentially assume a liquid supply posture in which each shelf surface is raised so that the communication portion is located at the bottom and liquid flows evenly into each shelf, an intermediate posture in which the cell culture vessel is rotated from that state so that the communication portion is located at the top and liquid in each shelf space is separated, and a culture posture in which the shelf surface of each shelf is subsequently rotated so that it is horizontal and liquid is distributed approximately evenly over each shelf surface.

6. The cell culture device according to any one of claims 3 to 6, wherein the drive mechanism comprises a first drive unit that rotates the cell culture vessel around a first axis, a second drive unit that rotates the cell culture vessel around a second axis perpendicular to the first axis, and a control unit that controls these drive units, wherein the first drive unit is located in a direction along the first axis, and the second drive unit is disposed in the vicinity of the first drive unit and is connected to the second axis via a transmission unit that transmits power along a direction parallel to the first axis.

7. The cell culture device according to any one of claims 3 to 6, further comprising an automatic winding mechanism for winding up excess length of a supply / discharge tube while applying a constant tension to the supply / discharge tube for a cell culture vessel that changes its posture while the supply / discharge tube is connected.

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