WO2021084737A1

WO2021084737A1 - Cell culture device

Info

Publication number: WO2021084737A1
Application number: PCT/JP2019/043036
Authority: WO
Inventors: 力三宅; 叶井　正樹
Original assignee: 株式会社島津製作所
Priority date: 2019-11-01
Filing date: 2019-11-01
Publication date: 2021-05-06
Also published as: JP7327501B2; JPWO2021084737A1

Abstract

This cell culture apparatus (100) comprises: a reading unit (137) that reads identification information (1a) attached to a cell culture device (1); and a control unit (140) that performs control regarding the movement of a culture chamber (10) due to a liquid feed gas (101) on the basis of the identification information read by the reading unit.

Description

Cell culture device

The present invention relates to a cell culture device, and more particularly to a cell culture device that transfers liquid between a plurality of culture chambers.

Conventionally, in cell culture, a technique of transferring a liquid between a plurality of culture chambers is known. Such techniques are disclosed, for example, in WO 2016/158233.

The International Publication No. 2016/158233 discloses a cell culture device (Body-on-a-Chip unit). According to the above-mentioned International Publication No. 2016/158233, the Body-on-a-chip system uses cultured cells as an organ model, and by connecting and culturing a plurality of organ models, in vivo in which a plurality of organs are involved. It is a method of reconstructing the phenomenon of the above in vitro. The Body-on-a-chip system is said to have potential as an alternative to animal experiments that can perform pharmacokinetic analysis that normally requires animal experiments in vitro, especially in cultured human-derived cells. If the analysis can be evaluated in vitro, it is expected that data with good correlation with clinical trials can be obtained.

The cell culture device of the above-mentioned International Publication No. 2016/158233 is composed of a total of 8 units, with a double culture chamber in which two culture chambers serving as an organ model are connected as one unit. The culture chambers constituting one unit are connected by a flow path. By sequentially pressurizing with a pneumatic pipe (pressurizing line), the culture solution can be circulated between each organ model culture chamber. In the above International Publication No. 2016/158233, the inside of the first culture chamber is pressurized through the first air filter, and the second culture chamber is opened to atmospheric pressure through the second air filter. The medium is sent from the first downstream port to the second culture chamber through the first communication channel.

International Publication No. 2016/158233

Here, it is considered that cell culture devices such as those disclosed in International Publication No. 2016/158233 may have different specifications (channels, etc.) from each other due to differences in organ models for which pharmacokinetic analysis is performed, for example. Be done. When automating cell culture, the user needs to make initial settings according to the cell culture device to be used, but it is complicated to make initial settings for each cell culture device. Therefore, even when cell culture devices (cell culture devices) having different specifications are used, it is possible to perform control suitable for the cell culture device (cell culture device) to be used while eliminating the complexity of initial setting. Is desired.

The present invention has been made to solve the above-mentioned problems, and one object of the present invention is to eliminate the complexity of initial setting even when cell culture devices having different specifications are used. At the same time, it is to provide a cell culture apparatus capable of performing control suitable for the cell culture device to be used.

In order to achieve the above object, the cell culture apparatus in one aspect of the present invention includes an installation unit in which a cell culture device having a plurality of culture chambers including a first culture chamber and a second culture chamber is installed, and a first. The liquid feed gas supply unit that supplies the liquid feed gas for moving the culture solution between the culture chamber and the second culture chamber, and the cell culture device installed in the installation unit and the liquid feed gas supply unit can be attached and detached. Based on the connecting gas pipe, the reading unit that reads the identification information attached to the cell culture device, and the identification information read by the reading unit, the culture solution between the first culture chamber and the second culture chamber using the liquid feed gas. It is provided with a control unit that controls the movement of the.

According to the present invention, as described above, the reading unit that reads the identification information attached to the cell culture device and the control unit that controls the cell culture device based on the identification information read by the reading unit. Provide. This relates to the movement of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas, which is suitable for the cell culture device to be used, while saving the trouble of performing the initial setting operation according to the cell culture device used by the user. Control can be performed. As a result, even when cell culture devices having different specifications are used, it is possible to perform control suitable for the cell culture device to be used while eliminating the complexity of initial setting.

It is a schematic diagram which showed the whole structure of the cell culture apparatus. It is a flow chart for demonstrating the cell culture method. It is a disassembled perspective view which showed the specific structural example of the cell culture device. It is a perspective view which showed the cell culture device in an assembled state. It is a figure which showed the circulation group containing two culture chambers. It is a figure which showed the connection part and the gas passage of a cell culture device. It is a vertical cross-sectional view which showed the connection part and the gas passage of a cell culture device. It is a figure which showed the operation procedure (A) and (B) of the circulation culture by the circulation group including two culture chambers. It is a figure which showed the operation procedure (A)-(D) of the circulation culture by the circulation group including four culture chambers. It is a schematic front view which showed the specific example of the cell culture apparatus. It is a top view which showed the structural example in the accommodating part of the cell culture apparatus shown in FIG. It is a schematic perspective view which showed the structural example of the installation part. It is a schematic diagram for demonstrating the connection between a cell culture device and a connector mechanism. It is a schematic top view of the cover member for demonstrating the structural example of the lid opening and closing mechanism. It is a schematic side view of the cell culture device for demonstrating the structural example of the lid opening and closing mechanism. It is a schematic diagram which showed the structural example of the liquid feed gas supply part. It is a schematic diagram which showed an example of the setting screen of the position of a culture chamber. It is a schematic diagram which showed an example of the setting screen of the movement of a culture solution between culture chambers. It is a schematic diagram which showed an example of the setting screen of the protocol of a cell culture device. It is a schematic diagram for demonstrating the reading of the identification information of consumables and chemicals. It is a flow chart which showed an example of the setting operation of a cell culture apparatus. It is a flow chart which showed an example of the processing operation of a cell culture apparatus. It is a flow chart which showed the processing of the sampling operation of FIG. 22. It is a flow chart which showed the process of the culture solution exchange operation of FIG. 22. It is a flow chart which showed the process of the reagent addition operation of FIG.

Hereinafter, embodiments embodying the present invention will be described with reference to the drawings.

The cell culture apparatus 100 and the cell culture method according to one embodiment will be described with reference to FIGS. 1 and 2.

(Cell culture device)
As shown in FIG. 1, the cell culture device 100 is a device that cultures cells 90 arranged in each culture chamber 10 by using a cell culture device 1 having a plurality of culture chambers 10. The cell culture device 100 is configured to automate at least a part of the cell culture process using the cell culture device 1. The culture chamber 10 is an example of the "first culture chamber" and the "second culture chamber" in the claims.

In the example shown in FIG. 1, the cell culture apparatus 100 includes an installation unit 110, a liquid feed gas supply unit 120, a gas pipe 130, a reading unit 137, and a control unit 140.

A cell culture device 1 having a plurality of culture chambers 10 is installed in the installation unit 110. The installation unit 110 is configured to be able to hold the cell culture device 1 in a predetermined position. The installation unit 110 has a contact surface that contacts at least a part of the cell culture device 1 and holds the cell culture device 1. The installation unit 110 has a mounting surface that supports the cell culture device 1 in contact with, for example, the lower surface of the cell culture device 1. The installation unit 110 has, for example, a pair of pressing surfaces that sandwich both side surfaces of the cell culture device 1 and support the cell culture device 1. The installation unit 110 has, for example, an engaging portion configured to be able to engage and disengage with a part of the cell culture device 1, and in the engaged state, the cell culture device 1 is fixedly held and engaged. Is released so that the cell culture device 1 can be removed.

The cell culture device 1 has a plurality of culture chambers 10. The culture chamber 10 is a storage space formed so as to store cells 90 and a liquid. The liquid is, for example, a culture medium of cells 90. The cells 90 are placed in the culture chamber 10, and the placed cells 90 are immersed in the culture medium. The cell culture device 1 is configured to enable perfusion culture in which a liquid (culture solution) is sent between a plurality of culture chambers 10. Therefore, the cell culture device 1 has a flow path 20 that fluidly connects between the plurality of culture chambers 10. In addition, perfusion is to pour a liquid such as a drug solution into an organ, a tissue, or a cell. In the present specification, "perfusion culture" refers to culturing by circulating the culture solution in the culture chamber 10 in the cell culture device 1, and the culture solution is statically cultured in the culture chamber 10. Distinguished from culture.

Identification information 1a for identifying the cell culture device 1 is attached to the cell culture device 1. One or more identification information 1a may be attached to the cell culture device 1. The identification information 1a is not particularly limited, but may be, for example, a one-dimensional code such as a barcode, a two-dimensional code such as a QR code (registered trademark), an RFID (Radio Frequency IDentifier) tag, or the like. Further, the identification information 1a may be, for example, a label affixed to the surface of the cell culture device 1.

The liquid feed gas supply unit 120 is configured to supply the liquid feed gas 101 for moving the culture liquid between the culture chambers 10. The liquid feed gas 101 is not particularly limited, but may be, for example, an atmosphere (air), a prepared gas different from the atmosphere, or the like.

The liquid feed gas supply unit 120 includes, for example, a gas source capable of supplying the atmosphere. The gas source is, for example, a pump that compresses and sends the atmosphere. The liquid feed gas supply unit 120 can supply the liquid feed gas 101 of the gas source, which is the atmosphere, to the cell culture device 1 via the gas pipe 130. The liquid feed gas supply unit 120 includes, for example, a gas source that stores a pre-prepared liquid feed gas 101. The gas source is, for example, a gas cylinder. The liquid feed gas supply unit 120 can supply the liquid feed gas 101 of the gas source, which is the adjusting gas, to the cell culture device 1 via the gas pipe 130. The liquid feed gas supply unit 120 is configured to prepare the liquid feed gas 101 from, for example, the component gas that is a component of the liquid feed gas 101. In this case, the liquid feed gas supply unit 120 includes one or more gas sources that store the component gas. The liquid feed gas supply unit 120 prepares the liquid feed gas 101 by, for example, mixing a plurality of component gases at a predetermined concentration ratio. The liquid feed gas supply unit 120 prepares the liquid feed gas 101 by, for example, mixing air taken in from the outside with one or a plurality of component gases at a predetermined concentration ratio. The liquid feed gas supply unit 120 can supply the prepared liquid feed gas 101 to the cell culture device 1 via the gas pipe 130. The liquid feed gas supply unit 120 is configured to be able to individually supply the liquid feed gas 101 to the plurality of culture chambers 10 via the gas pipe 130.

The gas pipe 130 is provided so as to detachably connect the cell culture device 1 installed in the installation unit 110 and the liquid feed gas supply unit 120. The gas pipe 130 is a tubular member having a first end and a second end opposite to the first end, and the first end and the second end are open, respectively. The first end of the gas pipe 130 is connected to the cell culture device 1, and the second end is connected to the liquid feed gas supply unit 120. As a result, the gas pipe 130 receives the liquid feed gas 101 from the liquid feed gas supply unit 120 through the opening at the second end and sends it out to the cell culture device 1 through the opening at the first end.

The gas pipe 130 can be individually connected to each of the plurality of culture chambers 10 so that the liquid feed gas 101 can be individually supplied to the plurality of culture chambers 10 of the cell culture device 1. In the example of FIG. 1, two gas pipes 130 are separately connected to the two culture chambers 10.

The reading unit 137 is configured to read the identification information 1a attached to the cell culture device 1. The reading unit 137 is not particularly limited, but can be a one-dimensional code reader when the identification information 1a is a one-dimensional code, and can be a two-dimensional code reader when the identification information 1a is a two-dimensional code. If the information 1a is an RFID tag, it can be an RFID reader. Further, the reading unit 137 may be, for example, a small terminal that can be grasped and used by the user.

The control unit 140 is composed of a computer (controller) that controls the operation of each unit of the cell culture device 100. The control unit 140 is electrically connected to, for example, the liquid feed gas supply unit 120, and controls the supply of the liquid feed gas 101 to the plurality of culture chambers 10. The control unit 140 controls perfusion to supply the liquid in the culture chamber 10 to another culture chamber 10 by supplying the liquid feed gas 101 to each culture chamber 10. That is, in the present embodiment, the control unit 140 supplies the liquid feed gas 101 to at least one of the plurality of culture chambers 10 via at least one flow path 20 connecting the plurality of culture chambers 10. It is configured to control the movement of the liquid in the culture chamber 10 to another culture chamber 10.

Specifically, the control unit 140 controls to pressurize and supply the liquid feed gas 101 to the culture chamber 10 which is the transfer source of the liquid. The control unit 140 controls to open the culture chamber 10 to which the liquid is moved to the outside or to supply a negative pressure to the culture chamber 10 to which the liquid is moved. As a result, the culture solution contained in the culture chamber 10 of the transfer source is pushed out of the culture chamber 10 by the supplied liquid feed gas 101, and moves to the culture chamber 10 of the transfer destination via the flow path 20. In the culture chamber 10 at the destination, the liquid feed gas 101 in the culture chamber 10 is pushed out by the volume integral of the inflowing culture solution.

For example, when the liquid feed gas supply unit 120 can supply the liquid feed gas 101 to an arbitrary culture chamber 10, the control unit 140 moves the liquid by controlling the liquid feed gas supply unit 120. In the example of FIG. 1, the cell culture device 100 includes a pressure control mechanism 135 provided in the middle of the gas pipe 130. The pressure control mechanism 135 includes, for example, a pressure control valve or a pressure regulator capable of controlling the pressure of the liquid feed gas 101 supplied from the liquid feed gas supply unit 120. The control unit 140 controls the pressure control mechanism 135 so as to move the liquid by pressurizing the inside of the culture chamber 10 with the liquid feed gas 101 when the liquid moves. That is, by controlling the pressure of the liquid feed gas 101 by the pressure control mechanism 135, the control unit 140 makes the pressure in the culture chamber 10 as the movement source higher than the pressure in the culture chamber 10 as the movement destination. As a result, the culture solution moves due to the pressure difference.

Further, in the example of FIG. 1, the cell culture device 100 includes a leak pipe 131 that is connected to the cell culture device 1 and discharges gas in the culture chamber 10, and a valve 132 that opens and closes the leak pipe 131. The leak pipe 131 is provided separately from the gas pipe 130 and is connected to the cell culture device 1 installed in the installation unit 110. The valve 132 is always closed and can be switched to the open state by the control of the control unit 140. In the example of FIG. 1, the leak pipe 131 and the valve 132 are individually connected to the plurality of culture chambers 10. The control unit 140 opens the valve 132 of the leak pipe 131 connected to the culture chamber 10 to be moved. As a result, the liquid feed gas 101 extruded with the movement of the culture solution is discharged to the outside.

Further, the control unit 140 is configured to adjust the component concentration of the liquid feed gas 101 in the culture chamber 10 via the leak pipe 131 by controlling the opening and closing of the valve 132. The control unit 140 adjusts the component concentration of the liquid feed gas 101 in the culture chamber 10 so that the component concentration of the liquid feed gas 101 is maintained at a predetermined value suitable for cell culture.

In the example of FIG. 1, the cell culture apparatus 100 is configured to enable circulation culture in which a liquid is circulated between a plurality of culture chambers 10. That is, after moving the liquid from one culture chamber 10 to the other culture chamber 10, the control unit 140 changes the relationship between the movement source and the movement destination, and changes the culture chamber 10 to which the liquid feed gas 101 is supplied. .. As a result, the liquid is moved in the opposite direction from the other culture chamber 10 to the one culture chamber 10. The control unit 140 controls so that perfusion from one culture chamber 10 to the other culture chamber 10 and perfusion from the other culture chamber 10 to one culture chamber 10 are alternately and repeatedly performed. As a result, cell culture can be performed while circulating the culture solution among the plurality of culture chambers 10.

In FIG. 1, the cell culture device 1 includes two culture chambers 10, but as will be described later, the cell culture device 1 may include three or more culture chambers 10. In that case, when the liquid is sent (when the culture liquid is moved), the valve 132 of the leak pipe 131 of the culture chamber 10 to be moved is opened, and each culture chamber 10 other than the culture chamber 10 to be moved is externally connected. The liquid feed gas 101 may be supplied at a pressure higher than the pressure.

Here, in the cell culture device 100, cell culture devices 1 having different specifications may be used. That is, in the cell culture device 100, the cell culture device 1 having different specifications such as the moving pressure of the culture solution between the culture chambers 10, the position of the culture chamber 10, the number of culture chambers 10, and the flow path 20 is installed in the installation unit 110. May be installed. Therefore, in the present embodiment, the control unit 140 is configured to control the cell culture device 1 based on the identification information 1a read by the reading unit 137. Specifically, the control unit 140 controls the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101 based on the identification information 1a read by the reading unit 137. In this case, the control unit 140 controls to acquire information regarding the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101 based on the identification information 1a read by the reading unit 137. The information regarding the movement of the culture solution between the culture chambers 10 by the acquired liquid delivery gas 101 may be included in the identification information 1a or may be stored in the cell culture apparatus 100. When stored in the cell culture device 100, the information regarding the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101 is stored in a state associated with the identification information 1a of the cell culture device 1.

For example, the control unit 140 cultures between the culture chambers 10 by the liquid feed gas 101 as information regarding the movement of the culture liquid between the culture chambers 10 by the liquid feed gas 101 based on the identification information 1a read by the reading unit 137. Obtain information on the moving pressure of the liquid. In this case, the control unit 140 uses, for example, information on the moving pressure of the culture solution between the culture chambers 10 by the acquired liquid feed gas 101, or information on which the setting is changed by the user for cell culture. It is set as information on the moving pressure of the culture solution between the culture chambers 10 of the cell culture device 1. Then, the control unit 140 controls the liquid transfer between the culture chambers 10 by the liquid feed gas 101 based on the information of the moving pressure of the culture solution between the set culture chambers 10. Information on the moving pressure of the culture solution between the culture chambers 10 is represented by, for example, the pressure in the culture chamber 10 as the moving source (pressurized side) and the pressure in the culture chamber 10 as the moving destination (non-pressurized side). be able to.

Further, for example, the control unit 140 uses the liquid feeding gas 101 as information regarding the movement of the culture liquid between the culture chambers 10 by the liquid feeding gas 101 based on the identification information 1a read by the reading unit 137. Get information on the movement timing of the culture solution. In this case, the control unit 140 uses, for example, information on the movement timing of the culture solution between the culture chambers 10 by the acquired liquid delivery gas 101, or information in which the setting is changed by the user for cell culture. It is set as information on the movement timing of the culture solution between the culture chambers 10 of the cell culture device 1. Then, the control unit 140 controls the liquid feeding between the culture chambers 10 by the liquid feeding gas 101 based on the information of the movement timing of the culture liquid between the set culture chambers 10. Information on the movement timing of the culture solution between the culture chambers 10 can be expressed by, for example, the movement time of the culture solution between the culture chambers 10.

Further, in the present embodiment, the control unit 140 controls the aspirated material in the culture chamber 10 of the plurality of culture chambers 10 based on the identification information 1a read by the reading unit 137 as control for the cell culture device 1. Controls suction. Further, the control unit 140 controls the discharge of the discharged material into the culture chamber 10 based on the identification information 1a read by the reading unit 137 as the control regarding the cell culture device 1. In these cases, the control unit 140 controls to acquire information on the positions of the plurality of culture chambers 10 based on the identification information 1a read by the reading unit 137. The information on the positions of the plurality of culture chambers 10 to be acquired may be included in the identification information 1a or may be stored in the cell culture apparatus 100. When stored in the cell culture device 100, the information on the positions of the plurality of culture chambers 10 is stored in a state associated with the identification information 1a of the cell culture device 1.

In this case, the control unit 140 uses, for example, the acquired information on the positions of the plurality of culture chambers 10 or the information in which the setting is changed by the user for culturing the cell culture device 1 used for cell culture. It is set as information on the position of the chamber 10. Then, the control unit 140 controls the discharge of the liquid as the discharge to the culture chamber 10, the suction control of the liquid as the suction from the culture chamber 10, and the like based on the information on the position of the set culture chamber 10. .. Information on the position of each of the plurality of culture chambers 10 can be represented by, for example, triaxial coordinates (XYZ coordinates) orthogonal to each other.

(Cell culture method)
Next, the cell culture method of this embodiment will be described. The cell culture method of the present embodiment is a cell culture method using the cell culture device 1. The cell culture method is carried out by the cell culture device 100 described above.

As shown in FIG. 2, in the cell culture method, at least the step 301 of preparing the liquid feed gas 101 for carrying out the liquid transfer between the culture chambers 10 and the prepared liquid feed gas 101 are used at least in the plurality of culture chambers 10. It includes a step 302 of moving the liquid in the culture chamber 10 to the other culture chamber 10 via the flow path 20 by supplying one.

In step 301, for example, a pump for compressing and sending the atmosphere is started, a gas cylinder storing a pre-prepared liquid feed gas 101 is set in the liquid feed gas supply unit 120, or the liquid feed gas supply unit 120 is set. This is carried out by preparing the liquid feed gas 101.

In step 302, the liquid feed gas 101 is pressurized and supplied to the culture chamber 10 which is the transfer source of the liquid, and the culture chamber 10 which is the transfer destination of the liquid is opened to the outside, or the culture chamber 10 which is the transfer destination of the liquid is opened. It is carried out by supplying a negative pressure to the chamber. As a result, the culture solution contained in the culture chamber 10 of the transfer source is pushed out of the culture chamber 10 by the supplied liquid feed gas 101, and moves to the culture chamber 10 of the transfer destination via the flow path 20. In the culture chamber 10 at the destination, the liquid feed gas 101 in the culture chamber 10 is pushed out by the volume integral of the inflowing culture solution.

In the cell culture method, the above-mentioned circulation culture may be performed. That is, in the cell culture method, the plurality of culture chambers 10 are obtained by changing the culture chamber 10 for supplying the liquid feed gas 101 and carrying out a plurality of steps of moving the liquid in the culture chamber 10 to another culture chamber 10. Further may be provided with steps (steps 303 and 304) of circulating the liquid between them.

In step 302, after perfusion from one culture chamber 10 to the other culture chamber 10, it is determined in step 303 whether or not to end the cell culture. If the cell culture is not completed, the process proceeds to step 304, and the culture chamber 10 for supplying the liquid feed gas 101 is changed. That is, the other culture chamber 10, which was the transfer destination in step 302, is set as the target for supplying the liquid feed gas 101. Then, the process proceeds to step 302, and this time, perfusion from the other culture chamber 10 to the one culture chamber 10 is performed. As a result, the culture solution is circulated between the plurality of culture chambers 10. The circulation culture is carried out a plurality of times until it is determined in step 303 that the cell culture is completed. When it is time to end the cell culture, it is determined in step 303 that the cell culture is finished, and the circulation culture is completed.

(Specific example of cell culture device)
A specific example of the cell culture device 1 will be described with reference to FIGS. 3 to 9. Here, a cell culture device 1 having a larger number of culture chambers 10 of 3 or more is shown. In the example of FIG. 3, the cell culture device 1 includes 16 culture chambers 10.

The cell culture device 1 includes a cover member 30, a through-hole plate 40, a microchannel plate 50, and a base member 60. The plurality of culture chambers 10 are formed as a storage space surrounded by a cover member 30, a through-hole plate 40, and a microchannel plate 50.

The through-hole plate 40 is formed with through-holes 41 that penetrate the through-hole plate 40 in the thickness direction (vertical direction) side by side. Through holes 41 are provided as many as the number of culture chambers 10. The inner peripheral surface of the through hole 41 of the through hole plate 40 constitutes the side wall surface of the culture chamber 10.

A micro flow path plate 50 in which a flow path 20 is formed is arranged below the through hole plate 40. The micro flow path plate 50 is provided so as to be in close contact with the lower surface of the through hole plate 40. The microchannel plate 50 closes the opening on the lower surface side of the through hole 41 and constitutes the bottom surface of the culture chamber 10.

The cover member 30 is arranged on the upper side of the through hole plate 40. The cover member 30 is provided so as to be in close contact with the upper surface of the through hole plate 40. The cover member 30 closes the opening on the upper surface side of the through hole 41 and constitutes the upper surface of the culture chamber 10.

Although not shown in FIG. 3, the cover member 30 has a gas supply port 31 (see FIGS. 6 and 7) for connecting to the gas pipe 130 and a liquid feed flowing from the gas supply port 31. A gas passage 32 (see FIGS. 6 and 7) for introducing the gas 101 into the culture chamber 10 and a gas discharge port 36 (see FIGS. 6 and 7) are provided. The structure of the cover member 30 will be described later.

The base member 60 has a recess for installing a laminate of the cover member 30, the through-hole plate 40, and the microchannel plate 50, and is configured to support the laminate arranged in the recess. A locking tool 61 for fixing the laminated body is attached to the base member 60. As shown in FIG. 4, the cell culture device 1 as an assembly is configured by fixing the laminated body to the base member 60 by the locking tool 61.

The culture chambers 10 are arranged in a matrix along the A and B directions that are orthogonal to each other. In FIG. 3, four culture chambers 10 are arranged in the A direction and four in the B direction, and are arranged in a 4 × 4 matrix.

The microchannel plate 50 is provided with a well 51 for placing cells 90 and a channel 20. The well 51 is a recess formed on the upper surface of the microchannel plate 50. The flow path 20 fluidly connects one culture chamber 10 and another culture chamber 10 through the inside of the micro flow path plate 50. The flow path 20 is provided so as to connect the culture chambers 10 arranged in the A direction.

By changing the shape of the flow path 20 formed in the micro flow path plate 50, the connection relationship between the plurality of culture chambers 10 can be changed. FIG. 3 shows two types of

microchannel plates

50a and 50b having different shapes of the channel 20. The microchannel plate 50a forms a circulation group 71 in which the two culture chambers 10 are connected by the channel 20. The microchannel plate 50b forms a circulation group 72 in which the four culture chambers 10 are connected by the channel 20. By selecting either one of the

microchannel plates

50a and 50b to form the laminate, the cell culture device 1 provided with the culture chamber 10 having an arbitrary connection relationship can be constructed.

In the configuration example of FIG. 5, the flow path 20 formed in the micro flow path plate 50 has a backflow prevention mechanism 21 that allows liquid to pass in one direction, and each of the plurality of culture chambers 10 is connected in a loop. There is. The individual culture chambers 10 are connected in a loop by the flow path 20 to form a circulation group 71. Circulation culture can be performed in which the culture solution is circulated between the culture chambers 10 constituting the circulation group 71.

Specifically, an introduction port 52 and an outlet port 53 formed in the microchannel plate 50 are arranged in one culture chamber 10. A flow path 20b connected to the culture chamber 10 on the upstream side is connected to the introduction port 52. A flow path 20a connected to the culture chamber 10 on the downstream side is connected to the outlet 53. The backflow prevention mechanism 21 is, for example, a check valve provided at the introduction port 52. Thereby, in the circulation culture, it is possible to transfer the liquid between the plurality of culture chambers 10 only in one direction from the upstream side to the downstream side. Thereby, in the cell culture method of the present embodiment, in the step of circulating the liquid between the plurality of culture chambers 10, the liquid is circulated in one direction among the plurality of culture chambers 10.

In the configuration example of FIG. 5, a gas inflow prevention mechanism 22 is provided at the outlet 53. The gas inflow prevention mechanism 22 is, for example, a Laplace valve. Laplace valves allow liquids to pass, but not gases, under pressure conditions below a given pressure. The predetermined pressure is at least higher than the pressurizing pressure when moving the liquid. As a result, when the liquid moves, the gas of the culture chamber 10 such as the liquid feed gas 101 is prevented from flowing into another culture chamber 10 through the flow path 20.

In the case of the microchannel plate 50a shown in FIG. 3, the channel 20 is formed so that the two culture chambers 10 arranged in the A direction form a circulation group. Therefore, the 16 culture chambers 10 form 8 circulation groups.

In the case of the microchannel plate 50b of FIG. 3, the channel 20 is formed so that the four culture chambers 10 arranged in the A direction form a circulation group. Therefore, the 16 culture chambers 10 form 4 circulation groups.

In each of the micro

flow path plates

50a and 50b, the flow path 20 is formed so that the

circulation groups

71 and 72 having the same connection relationship are lined up in the B direction. Therefore, when moving the liquid, the liquid feeding operation of each circulation group arranged in the B direction (moving operation of the culture liquid) is performed only by pressurizing and supplying the liquid feeding gas 101 to the four culture chambers 10 arranged in the B direction at once. ) Can be implemented together.

Therefore, in the examples shown in FIGS. 6 and 7, the gas supply port is provided so that the liquid feed gas 101 can be collectively pressurized and supplied to a plurality of culture chambers 10 arranged in the B direction at the same position in the A direction. 31 and a gas passage 32 are provided.

Further, in the examples shown in FIGS. 6 and 7, a gas discharge port is provided so that the gas in the culture chamber 10 can be collectively discharged to a plurality of culture chambers 10 arranged in the B direction at the same position in the A direction. 36 is provided. Therefore, the cell culture device 1 includes a gas supply port 31 for supplying the liquid feed gas 101 to the culture chamber 10 and a gas discharge port 36 for discharging the gas in the culture chamber 10 to the valve 132.

Specifically, in the example shown in FIG. 7, the cover member 30 is arranged between the lid 33 on the upper surface side, the sealing plate 34 in contact with the through-hole plate 40, and the lid 33 and the sealing plate 34. Also includes an intermediate plate 35. A gas passage 32 is provided inside the intermediate plate 35. The lid 33 covers the upper surface of the intermediate plate 35 and seals the gas passage 32 from the upper surface side. The lid 33 constitutes the upper surface of the entire cell culture device 1. The sealing plate 34 seals between the through-hole plate 40 (culture chamber 10) and the intermediate plate 35 to prevent leakage from a portion other than the gas passage 32. The sealing plate 34 is formed with a through hole that penetrates the sealing plate 34 in the thickness direction at a position directly above the culture chamber 10. The gas passage 32 of the intermediate plate 35 and the culture chamber 10 are fluidly connected to each other through the through hole of the sealing plate 34.

In the example shown in FIG. 6, the gas supply port 31 and the gas discharge port 36 communicating with the same culture chamber 10 are provided so as to project laterally from the side surfaces opposite to each other of the cell culture device 1.

Specifically, the cell culture device 1 has four gas supply ports 31 arranged in the A direction, four gas discharge ports 36 arranged in the A direction, and four gas passages 32 arranged in the A direction. It is provided. A pair of a gas supply port 31 and a gas discharge port 36 arranged in the B direction communicate with each other through one gas passage 32 (see FIG. 7) extending in the B direction. Each gas passage 32 is connected in parallel to four culture chambers 10 arranged in the B direction. When the liquid feed gas 101 is supplied from any of the gas supply ports 31, the liquid feed gas 101 is collectively supplied to the four culture chambers 10 arranged in the B direction via the gas passage 32. Therefore, for the supply of the liquid feed gas 101, it is only necessary to select one of the four gas supply ports 31. Similarly, for gas discharge in the culture chamber 10, it is only necessary to select one of the four gas discharge ports 36.

As described above, the plurality of gas pipes 130 and the plurality of leak pipes 131 are the respective culture chambers 10 (individual culture chambers 10 included in the circulation group) interconnected via the flow path 20 in the cell culture device 1. ), It is provided so as to be individually connected. On the other hand, the plurality of gas pipes 130 and the plurality of leak pipes 131 are connected in parallel to the plurality of circulation groups arranged in the B direction via the gas passage 32.

A specific operation example in circulation culture will be explained.

FIG. 8 shows an operation example of the circulation group 71 composed of two

culture chambers

10a and 10b. The circulation group 71 circulates the liquid by repeating the operations (A) and (B). In (A), the liquid feed gas 101 is pressurized and supplied to the culture chamber 10a to open the culture chamber 10b to the outside. As a result, the culture solution in the culture chamber 10a moves to the culture chamber 10b via the flow path 20a. In (B), the culture chamber 10a is opened to the outside, and the liquid feed gas 101 is pressurized and supplied to the culture chamber 10b. As a result, the culture solution in the culture chamber 10b moves to the culture chamber 10a via the flow path 20b.

FIG. 9 shows an operation example of the circulation group 72 composed of four

culture chambers

10a, 10b, 10c and 10d. The circulation group 72 circulates the liquid by repeating the operations (A) to (D). In (A), the liquid feed gas 101 is pressurized and supplied to the culture chamber 10a to open the culture chamber 10b to the outside. As a result, the culture solution in the culture chamber 10a moves to the culture chamber 10b via the flow path 20a. In (B), the liquid feed gas 101 is pressurized and supplied to the culture chamber 10b to open the culture chamber 10c to the outside. As a result, the culture solution in the culture chamber 10b moves to the culture chamber 10c via the flow path 20b. In (C), the liquid feed gas 101 is supplied to the culture chamber 10c, and the culture chamber 10d is opened to the outside. As a result, the culture solution in the culture chamber 10c moves to the culture chamber 10d via the flow path 20c. In (D), the liquid feed gas 101 is supplied to the culture chamber 10d, and the culture chamber 10a is opened to the outside. As a result, the culture solution in the culture chamber 10d moves to the culture chamber 10a via the flow path 20d.

With such a configuration, circulation culture in which the culture solution is circulated between the plurality of culture chambers 10 connected by the flow path 20 is possible.

In the present embodiment, cells 90 differentiated into different types are arranged in the plurality of culture chambers 10. That is, cells 90 of different types are seeded in the wells 51 of the individual culture chambers 10 that form the circulation group. In the present embodiment, organ model cells 90 derived from different organs are arranged in the plurality of culture chambers 10.

The organ model cell 90 uses cells derived from the specific organ as cultured cells in order to simulate the intra-organ dynamics such as absorption, metabolism, and excretion in the specific organ in pharmacokinetic analysis and the like. Different organ model cells 90 are seeded in individual culture chambers 10 constituting the circulation groups (71, 72), and by performing circulation culture, in vivo phenomena involving a plurality of organs can be exhibited in the cell culture device 1. It is possible to simulate with.

The organ model cell 90 is a cell derived from an organ such as the heart, lung, liver, kidney, and intestine. The organ model cell 90 can be a cell derived from an organ other than these. The cell 90 seeded in the culture chamber 10 may be a cell that models a plurality of different sites of a specific organ, instead of modeling a specific organ. The cell 90 may be, for example, a specific cancer cell. For example, in the example of FIG. 8, the liver organ model cells 90 are seeded in the culture chamber 10a, and the cancer cells 90 are seeded in the culture chamber 10b. For example, in the example of FIG. 9, lung organ model cells 90 are in the culture chamber 10a, liver organ model cells 90 are in the culture chamber 10b, liver organ model cells 90 are in the culture chamber 10c, and kidney organ model cells 90 are in the culture chamber 10d. , Are sown.

(Specific example of cell culture device)
A specific example of the cell culture apparatus 100 will be described with reference to FIGS. 10 and 11.

In the examples of FIGS. 10 and 11, the cell culture device 100 is configured to be able to automatically perform various operations associated with cell culture using the cell culture device 1. Specifically, in addition to the circulation culture of cells, the cell culture device 100 can dispense the liquid into the culture chamber 10, suck the liquid in the culture chamber 10, and control the temperature of the cell culture device 1.

In the example of FIG. 10, the cell culture device 100 includes a storage unit 102 that forms a working space in a clean air environment, and a base unit 103 on which the storage unit 102 is installed. The operations associated with cell culture are performed inside the containment unit 102. The cell culture device 100 includes a liquid feeding mechanism 150 that feeds a liquid. In the present specification, "clean air" is air from which impurities such as dust and microorganisms contained in the atmosphere have been removed.

The accommodating unit 102 is configured to accommodate the installation unit 110 and the liquid feeding mechanism 150 in a clean air environment. The accommodating portion 102 has a box shape that covers the upper surface of the base portion 103. Clean air is supplied into the accommodating portion 102 from an air intake portion (not shown) provided with an air filter. The air filter is, for example, a HEPA (High Efficiency Particulate Air) filter. The accommodating portion 102 is configured to thereby maintain the internal space in a clean air environment.

As shown in FIG. 11, an installation unit 110, a liquid feeding mechanism 150, a container cooling unit 160, a heating unit 170, and a container transfer mechanism 180 are arranged in the accommodating unit 102.

The cell culture device 1 is placed on the installation unit 110. In the configuration example of FIG. 11, two installation units 110 are shown for convenience of illustration. The number of installation units 110 is not limited to two, and may be one or three or more. The specific configuration of the installation unit 110 will be described later.

The liquid feeding mechanism 150 sucks and discharges the liquid under the control of the control unit 140. The control unit 140 is at least one of a process of sucking the liquid contained in any of the plurality of culture chambers 10 and a process of discharging the liquid into any of the plurality of culture chambers 10 at a preset timing. The liquid feeding mechanism 150 is controlled so as to carry out.

As shown in FIG. 10, the liquid feeding mechanism 150 includes a dispensing mechanism 151 that sucks and discharges a liquid, and a moving mechanism 152 that moves the dispensing mechanism 151 between a discharge position or a suction position and an installation unit 110. And include. The dispensing mechanism 151 is an example of the "first mechanism" in the claims. Further, the moving mechanism 152 is an example of the "second mechanism" in the claims.

The dispensing mechanism 151 is provided with a suction pipe 153 to which a pipette tip 191 can be attached at the tip, and is configured to be able to supply positive pressure and negative pressure to the suction pipe 153. The accommodating unit 102 is provided with a chip rack installation unit 190 on which a chip rack 192 holding a plurality of disposable pipette tips 191 is placed. In the example of FIG. 11, eight chip racks 192 can be installed in the chip rack installation unit 190, and only one chip rack 192 is shown. The accommodating portion 102 is provided with a waste port 104 for disposing of the used pipette tip 191 and the like, and a waste liquid port 105 for disposing of the sucked liquid.

As shown in FIG. 10, the moving mechanism 152 is provided near the ceiling of the accommodating portion 102, and supports the dispensing mechanism 151 in a suspended state. The moving mechanism 152 is configured so that the dispensing mechanism 151 can be moved in the vertical direction and the horizontal direction in the accommodating portion 102. The moving mechanism 152 is, for example, a gantry-type Cartesian robot that can move in three orthogonal axial directions. In FIG. 11, the moving mechanism 152 moves the dispensing mechanism 151 to the upper positions of the installation unit 110, the container cooling unit 160, the heating unit 170, the chip rack installation unit 190, the waste port 104, and the waste liquid port 105 described later. Can be moved.

In the example of FIG. 11, the container cold insulation unit 160 is configured to maintain and store the liquid container at a predetermined storage temperature. The container cold insulation unit 160 is provided with a plurality of container installation portions so that various liquid containers can be installed.

Specifically, the container cold insulation unit 160 includes a container installation unit 161 in which the sample container 161a can be installed. The container cold insulation unit 160 includes a container installation unit 162 in which the reagent container 162a can be installed. The container cold insulation unit 160 includes a container installation unit 163 in which the culture solution container 163a can be installed.

The container installation unit 161, the container installation unit 162, and the container installation unit 163 are all configured so that a container rack holding a plurality of containers can be placed. In the example of FIG. 11, the container installation unit 161 can install eight container racks 161b capable of holding the sample container 161a. In the example of FIG. 11, the sample container 161a is, for example, a well plate (also referred to as a microplate) in which a plurality of accommodating recesses are integrally formed, but one sample tube is held in the container rack 161b (see FIG. 12). Can be. In the example of FIG. 11, the container installation unit 162 can install one container rack 162b capable of holding four reagent containers 162a. In the example of FIG. 11, the container installation unit 163 can install one container rack 163b capable of holding 20 culture solution containers 163a. The number of containers or container racks that can be installed in the container installation section is not particularly limited.

The sample container 161a is a container for accommodating the culture solution (sample) obtained (sampled) from the culture chamber 10 when analyzing the culture solution in the culture chamber 10 for pharmacokinetic analysis or the like. The reagent container 162a is a container containing a drug (reagent) to be evaluated in pharmacokinetic analysis or the like. The culture solution container 163a is a container containing an unused culture solution used for cell culture. The unused culture medium is used for exchanging or replenishing the culture medium in the culture chamber 10 in the process of cell culture treatment.

The container cold insulation unit 160 includes a second temperature control mechanism 164 that adjusts the containers installed in the container installation unit 161, the container installation unit 162, and the container installation unit 163 to a storage temperature lower than that in the culture chamber 10. The second temperature control mechanism 164 includes, for example, a Peltier element, and maintains the container installation unit at the storage temperature under the control of the control unit 140. The container cold storage unit 160, for example, refrigerates the container at 10 ° C. or lower. The storage temperature is, for example, 4 ° C.

The heating unit 170 is configured to preheat the liquid to be dispensed when the liquid in the container stored cold in the container cold storage unit 160 is dispensed into the culture chamber 10.

Specifically, the heating unit 170 includes a third temperature control mechanism 171. The third temperature control mechanism 171 is configured so that a container for containing the liquid can be installed. In the example of FIG. 11, the third temperature control mechanism 171 has 12 holding holes 172 for receiving and holding the container. In the holding hole 172, a reagent container 162a stored in the container cold insulation unit 160, a culture solution container 163a, and a disposable container 173 for subdividing a required amount of liquid from these containers can be installed. The third temperature control mechanism 171 adjusts the temperature of the liquid in the container installed in the holding hole 172 so as to be close to the temperature in the culture chamber 10. The third temperature control mechanism 171 is composed of, for example, a heater such as a heating wire, a Peltier element, or the like. The third temperature control mechanism 171 is controlled based on the control of the control unit 140 so as to match, for example, the set temperature in the culture chamber 10 (the installation temperature of the first temperature control mechanism 112 described later).

The container transfer mechanism 180 is configured to be able to transfer a liquid container in the accommodating portion 102. The container transfer mechanism 180 can take out the container from the container cold insulation unit 160 and transfer it to the heating unit 170. The container transfer mechanism 180 can take out the container installed in the heating unit 170 and transfer it to the container cold insulation unit 160. The container transfer mechanism 180 can transfer the used container in the container cooling unit 160 or the heating unit 170 to the disposal port 104 and dispose of it.

As shown in FIG. 10, the container transfer mechanism 180 includes a gripping mechanism 181 that grips the container so that it can be released, and a moving mechanism 182 that moves the gripping mechanism 181. The gripping mechanism 181 is composed of, for example, a hand mechanism that can operate to hold the container, and a chuck mechanism such as a vacuum chuck or a magnetic chuck. The moving mechanism 182 is configured to be able to move the gripping mechanism 181 in the vertical direction and the horizontal direction in the accommodating portion 102. The moving mechanism 182 is, for example, a gantry-type Cartesian robot that can move in three orthogonal axial directions.

In the example of FIG. 10, the base unit 103 houses the liquid feed gas supply unit 120 and the control unit 140. Further, a waste storage 106 and a waste liquid tank 107 are installed on the base 103.

The liquid feed gas supply unit 120 includes a pressure control mechanism 135 and a flow rate control mechanism 136.

The control unit 140 includes a processor 141 such as a CPU and a storage unit 142 which is a volatile memory. The processor 141 functions as a control unit that controls each unit of the cell culture apparatus 100 by executing a program stored in the storage unit 142. The storage unit 142 stores the setting information of the cell culture device 100.

The setting information includes, for example, the device information of the cell culture device 1. The device information includes information such as the structure of the cell culture device 1, for example, the position and number of culture chambers 10. The setting information includes, for example, consumables information. The consumables information includes information such as the type and holding position of the container (sample container 161a) stored in the container cold insulation unit 160, the position of the pipette tip 191 (or tip rack 192), and the like. The setting information includes, for example, liquid information. The liquid information includes information such as the type, contents, amount of liquid to be contained, and holding position of the reagent container 162a and the culture solution container 163a.

The setting information includes, for example, schedule information. The schedule information is information that sets the timing of the operation associated with the cell culture by the cell culture device 100. The schedule information is set, for example, the timing of sampling the culture solution, the timing of reagent dispensing, the timing of exchanging the culture solution, and the like. The control unit 140 executes these operations at the timing set in the schedule information.

The waste storage 106 is connected to a waste port 104 (see FIG. 11) formed on the upper surface of the base portion 103, and is configured to store the waste put into the waste port 104. The waste is a used pipette tip 191 and a used liquid container.

The waste liquid tank 107 is connected to a waste liquid port 105 (see FIG. 11) formed on the upper surface of the base portion 103, and is configured to store the liquid charged into the waste liquid port 105. The waste liquid is a surplus amount of liquid sucked by the dispensing mechanism 151, a used culture liquid sucked from the culture chamber 10 when exchanging the culture liquid, and other liquids in the container that are no longer needed.

(Installation part)
As shown in FIG. 12, a cell culture device 1 having a plurality of culture chambers 10 is installed in the installation unit 110. The installation unit 110 detachably supports the cell culture device 1. In the example of FIG. 12, the installation unit 110 includes a mounting surface 111 on which the cell culture device 1 is placed, and supports the lower surface of the cell culture device 1. The installation unit 110 may have a slot corresponding to the outer shape of the cell culture device 1, for example, and the cell culture device 1 may be inserted into the slot.

In the example of FIG. 12, the cell culture device 100 is configured to adjust the temperature of the cell culture device 1 installed in the installation unit 110 so as to maintain the temperature suitable for cell culture.

That is, the cell culture device 100 includes a first temperature control mechanism 112 that controls the temperature of the cell culture device 1 installed in the installation unit 110. The first temperature control mechanism 112 is configured to be maintained at a predetermined set temperature under the control of the control unit 140. The set temperature is, for example, 36 ° C to 37 ° C. The first temperature control mechanism 112 includes, for example, a heater and generates heat to heat the cell culture device 1. An example of the heater is a cartridge heater in which a heating wire is housed in a protective tube, but the heater is not particularly limited.

The first temperature control mechanism 112 is provided so as to be adjacent to any one or more of the lower surface, the upper surface, and the side surface of the cell culture device 1. In the example of FIG. 12, the first temperature control mechanism 112 is provided so that the cell culture device 1 is adjacent to the lower surface. That is, the upper surface of the first temperature control mechanism 112 constitutes the mounting surface 111 of the installation portion 110.

Further, in the example of FIG. 12, in the cell culture device 100, the cell culture device 1 is installed in the installation unit 110, so that the liquid feed gas supply unit 120 and the cell culture device 1 are connected via the gas pipe 130. It is configured to. That is, the user only needs to set the cell culture device 1 in the installation unit 110, and the connection between the cell culture device 1 and the gas pipe 130 is completed.

Specifically, the cell culture device 100 includes a connector mechanism 113 that is arranged in the installation unit 110 and is connected to a plurality of gas pipes 130. The connector mechanism 113 is configured to connect the plurality of gas pipes 130 and the corresponding plurality of culture chambers 10 by installing the cell culture device 1 in the installation unit 110.

In the example shown in FIG. 13, the cell culture device 1 is provided with a gas supply port 31 for connecting to the gas pipe 130. The connector mechanism 113 has an insertion port 114 into which the gas supply port 31 of the cell culture device 1 is inserted, and a valve 115. By inserting the gas supply port 31 into the insertion port 114, the culture chamber 10 of the cell culture device 1 and the gas pipe 130 inside the connector mechanism 113 are connected. The cell culture device 1 is provided with four gas supply ports 31. In this case, the connector mechanism 113 is provided with four insertion ports 114 (see FIG. 12) and four valves 115 corresponding to the four gas supply ports 31. When the cell culture device 1 is set on the mounting surface 111 of the installation unit 110, the four gas supply ports 31 are inserted into the corresponding four insertion ports 114, respectively.

Further, in the example of FIG. 13, the cell culture device 100 includes a discharge mechanism 116 that is detachably attached to the gas discharge port 36 of the cell culture device 1. The discharge mechanism 116 includes four leak pipes 131 and four valves 132. The discharge mechanism 116 has four insertion ports 117 into which the gas discharge ports 36 are inserted, corresponding to the four gas discharge ports 36 of the cell culture device 1. The four leak pipes 131 and the four valves 132 are provided separately for the four insertion ports 114. The leak pipe 131 is open to the space inside the accommodating portion 102 outside the discharge mechanism 116.

The four valves 115 and the four valves 132 can be opened and closed under the control of the control unit 140. As a result, it is possible to arbitrarily perform the feeding of the culture solution (that is, circulation culture) under the control of the control unit 140.

The connector mechanism 113 is fluidly connected to the liquid feed gas supply unit 120 via the gas pipe 130. Further, in the example of FIG. 13, a leak pipe 131 connected to the gas pipe 130 and a valve 132 for opening and closing the leak pipe 131 are provided in the connector mechanism 113. The valve 132 is, for example, a three-way valve. The valve 132 is controlled by the control unit 140, for example, in a state where the insertion port 114 and the liquid feed gas supply unit 120 are connected, a state in which the insertion port 114 and the leak pipe 131 are connected, an insertion port 114, and a liquid feed supply unit. It is possible to switch between a state in which the gas supply unit 120 and the leak pipe 131 are cut off from each other (fully closed state). The leak pipe 131 is open to the space inside the accommodating portion 102 outside the connector mechanism 113.

The leak pipe 131 and the valve 132 are separately provided for the four insertion ports 114. Therefore, the process of supplying the liquid feed gas 101 from the liquid feed gas supply unit 120 to the four gas supply ports 31 of the cell culture device 1 and the gas supply port 31 to the outside via the leak pipe 131 are individually performed. It is possible to switch between the process of opening and the process of opening. Thereby, the liquid feeding (that is, circulation culture) of the culture solution shown in FIGS. 8 and 9 can be arbitrarily performed under the control of the control unit 140.

Further, in the example shown in FIG. 12, the cell culture device 100 can automatically open and close the lid 33 of the cell culture device 1 so that the dispensing mechanism 151 can access the inside of the culture chamber 10 in the cell culture device 1. It is configured.

Specifically, the cell culture device 100 includes a lid opening / closing mechanism 210 that opens / closes the lid 33 that seals the culture chamber 10 of the cell culture device 1. The lid opening / closing mechanism 210 is configured to open / close the lid 33 by moving the lid 33 of the cell culture device 1 placed on the installation unit 110. The lid opening / closing mechanism 210 includes an actuator that is a drive source for the lid 33. The actuator can be an electric motor, an air cylinder, or the like. The control unit 140 controls the lid opening / closing mechanism 210, and also controls the moving mechanism 152 so that the dispensing mechanism 151 accesses the culture chamber 10 in which the lid 33 is opened by the lid opening / closing mechanism 210.

For example, in the cell culture device 1 shown in FIGS. 3 and 4, the upper portions of the 16 culture chambers 10 are collectively covered by the lid 33 of the cover member 30. The lid opening / closing mechanism 210, for example, separates the lid 33 shown in FIG. 4 from the cover member 30, and opens the upper portions of the 16 culture chambers 10 together. After the suction operation and the discharge operation are performed by the dispensing mechanism 151, the lid opening / closing mechanism 210 places the lid 33 separated from the cover member 30 again on the intermediate plate 35 to seal the culture chamber 10.

The plurality of culture chambers 10 need not be sealed by one lid 33, and as shown in FIG. 12, the plurality of lids 33 may separately cover a group of several culture chambers 10. In the example shown in FIG. 12, the cell culture device 1 is provided with four lids 33 covering the four culture chambers 10 arranged in the A direction in the B direction. That is, the two circulation groups 71 composed of the two culture chambers 10 shown in FIG. 3 or the one circulation group 72 composed of the four culture chambers 10 shown in FIG. 3 can be opened and closed as a unit. Is provided with a lid 33. The lid opening / closing mechanism 210 is configured so that the four lids 33 can be opened / closed individually. As a result, only the culture chambers 10 that form a specific circulation group can be opened and closed individually.

For example, the same number of lids 33 as the number of culture chambers 10 may be provided for each culture chamber 10. In this case, the lid opening / closing mechanism 210 may be configured to open / close the individual lids 33 separately. As a result, the culture chambers 10 can be opened and closed separately one by one.

The lid opening / closing mechanism 210 can move the lid 33 to the open position 401 and the closed position 402. With the lid 33 in the open position 401, the upper part of the culture chamber 10 is opened in the accommodating portion 102, and the dispensing mechanism 151 becomes accessible in the culture chamber 10. With the lid 33 in the closed position 402, the liquid feed gas 101 can be pressurized and supplied to the culture chamber 10 through the gas passage 32.

The dispensing mechanism 151 sucks the liquid from the container held in the heating unit 170, for example, and discharges the liquid into the culture chamber 10 of the cell culture device 1 installed in the installation unit 110. In this case, the suction position is the upper position of the container of the heating unit 170, and the discharge position is the upper position of the culture chamber 10. The dispensing mechanism 151 is configured to dispense, for example, the liquid in the reagent container 162a into the culture chamber 10.

The dispensing mechanism 151 sucks the liquid from, for example, the culture chamber 10 of the cell culture device 1 installed in the installation unit 110, and discharges the liquid into the container held in the container cold insulation unit 160 or to the waste liquid port 105. The dispensing mechanism 151 is configured to dispense, for example, the liquid in the culture chamber 10 into the sample container 161a. The dispensing mechanism 151 is configured to exchange the culture solution by, for example, sucking the culture solution in the culture chamber 10 and dispensing the liquid in the culture solution container 163a into the culture chamber 10. There is.

(Specific example of lid opening / closing mechanism)
In the configuration example shown in FIGS. 14 and 15, the lid opening / closing mechanism 210 opens / closes the lid 33 by moving the regulating member 223 that regulates the movement of the lid 33.

In the example of FIG. 14, four lids 33 covering the four culture chambers 10 arranged in the A direction are provided in the B direction. In FIG. 14, the lid 33 is shown with hatching for convenience. As shown in FIG. 15, the lid 33 is attached to the intermediate plate 35 in a state of being rotatable via the hinge 221. The hinge 221 is provided with an urging member 222 that urges the lid 33 in the opening direction (the direction away from the intermediate plate 35). The urging member 222 is, for example, a torsion spring.

A regulation member 223 extending in the A direction is provided on the upper surface of the lid 33. As shown in FIG. 14, the regulating member 223 extends in the A direction to both outer sides of the lid 33, and both ends thereof are held by the guide portions 224 so as not to be movable in the vertical direction. The guide portion 224 holds the regulation member 223 movably in the B direction at the regulation position 225 on the upper surface of the lid 33 and the release position 226 which is separated from the lid 33 in the B direction.

As shown in FIG. 15, the lid opening / closing mechanism 210 includes an engaging portion 211 that engages with the regulation member 223 and a driving portion 212 that is an actuator that moves the engaging portion 211 in the B direction.

The lid opening / closing mechanism 210 moves the restricting member 223 from the restricting position 225 to the releasing position 226 by moving the engaging portion 211. As a result, the lid 33 is released from the regulation by the regulating member 223, and rotates to the open position 401 according to the urging force of the urging member 222.

The lid opening / closing mechanism 210 moves the regulation member 223 from the release position 226 to the regulation position 225 by moving the engaging portion 211. In the process of movement, the regulating member 223 rotates the lid 33 in the closing direction against the urging force of the urging member 222. When the regulating member 223 reaches the regulating position 225, the lid 33 moves to the closing position 402 in which the lid 33 is in close contact with the intermediate plate 35, and the upper part of the culture chamber 10 is sealed. The regulation member 223 regulates the movement of the lid 33 at the regulation position 225 so as not to be rotated by the urging force of the urging member 222.

(Liquid feed gas supply unit)
FIG. 16 shows a configuration example of the liquid feed gas supply unit 120. In the example of FIG. 16, the liquid feed gas supply unit 120 is connected to the cell culture device 1 via the gas pipe 130 connected to the connector mechanism 113.

In the configuration example of FIG. 16, the liquid feed gas supply unit 120 includes an air intake unit 121, a gas source 122 of a gas to be mixed such as carbon dioxide, and a mixing chamber 123 for mixing air and the gas to be mixed. , Is configured to prepare the feed gas 101.

The air intake unit 121 is connected to the mixing chamber 123 via a pipe. The air intake unit 121 takes in air from the outside of the cell culture device 100 and allows it to pass through an air filter to generate clean air. The air filter is, for example, a HEPA filter. The air intake unit 121 supplies the generated clean air to the mixing chamber 123.

The gas source 122 is a gas cylinder that stores the gas to be mixed at high pressure. The gas source 122 is connected to the mixing chamber 123 via a pipe. The gas source 122 supplies the gas to be mixed to the mixing chamber 123.

The mixing chamber 123 is a gas container having a volume capable of storing a predetermined amount of gas. The mixing chamber 123 is connected to the supply control unit 124. The mixing chamber 123 mixes the air (clean air) supplied from the air intake unit 121 and the gas to be mixed supplied from the gas source 122.

Water 123a is stored in the mixing chamber 123. The mixing chamber 123 is configured to discharge air and a gas to be mixed inside the water 123a. That is, the mixing chamber 123 is configured to humidify the supplied gas by bubbling the air and the gas to be mixed in the water 123a.

In the mixing chamber 123, a gas sensor 123b to be mixed that detects the concentration of the gas to be mixed in the gas, a pressure sensor 123c that detects the pressure of the gas in the mixing chamber 123, and the temperature of the gas in the mixing chamber 123 are detected. A temperature sensor 123d is provided. The control unit 140 controls the supply of air and the mixed gas to the mixing chamber 123 based on the detection signals of the mixed gas sensor 123b, the pressure sensor 123c, and the temperature sensor 123d. As a result, the liquid feed gas 101 having a saturated vapor pressure having a predetermined gas concentration to be mixed is prepared in the mixing chamber 123.

The supply control unit 124 is connected to the connector mechanism 113 via the gas pipe 130. The supply control unit 124 is configured to send the liquid feed gas 101 prepared in the mixing chamber 123 to the gas pipe 130 under the control of the control unit 140.

The supply control unit 124 can individually supply the liquid feed gas 101 to each gas supply port 31 of the cell culture device 1 via the connector mechanism 113. Therefore, the supply control unit 124 is provided with the same number of connection ports 125 as the gas supply port 31 (insertion port 114 of the connector mechanism 113), and one gas pipe 130 is connected to each connection port 125.

The supply control unit 124 includes a pressure control mechanism 135 including a pressure control valve or a pressure regulator, and a flow rate control mechanism 136 including a flow rate sensor and a flow rate control valve. The pressure control mechanism 135 and the flow rate control mechanism 136 are individually provided for each connection port 125. The control unit 140 controls the supply pressure and the gas flow rate of the liquid feed gas 101 to the individual gas pipes 130 by controlling the pressure control mechanism 135 and the flow rate control mechanism 136.

Each unit of the liquid feed gas supply unit 120 having the above configuration is controlled by the control unit 140 (see FIG. 10). The control unit 140 is configured to perform control (circulation culture) to circulate the liquid among the plurality of culture chambers 10 by sequentially switching the culture chambers 10 for supplying the liquid feed gas 101.

Further, the control unit 140 (see FIG. 10) controls the pressure control mechanism 135 so as to move the liquid by pressurizing the inside of the culture chamber 10 with the liquid feed gas 101 when the liquid (culture liquid) is moved, and culture. After moving the liquid in the chamber 10 to another culture chamber 10, by opening the valve 132, the inside of the culture chamber 10 at the movement source is moved under the atmosphere of the liquid feed gas 101 having a pressure lower than the pressurized pressure. The pressure control mechanism 135 is controlled so as to be. The pressure of the liquid feed gas 101 when moving the liquid is, for example, about 5 kPa (gauge pressure). The pressure of the liquid feed gas 101 after moving the liquid is, for example, atmospheric pressure (0 kPa in gauge pressure). When the liquid moves, the control unit 140 pressurizes and supplies the liquid feed gas 101 to the culture chamber 10, and then temporarily opens the culture chamber 10 to the atmosphere through the leak pipe 131 to bring the inside of the culture chamber 10 to atmospheric pressure. maintain.

When the liquid is moved, the control unit 140 (see FIG. 10) pressurizes the culture chamber 10 that is the source of the liquid movement by the liquid feeding gas 101 of the liquid feeding gas supply unit 120, and inside the culture chamber 10 that is the destination of the liquid movement. Is controlled to be opened to the outside by the leak pipe 131.

(Reading unit, display unit, operation unit)
In the example of FIG. 10, the reading unit 137, the display unit 138, and the operating unit 139 are electrically connected to the control unit 140. The reading unit 137 reads the identification information 1a (see FIG. 1) attached to the cell culture device 1 and transmits it to the control unit 140. The display unit 138 includes, for example, a liquid crystal monitor, and displays information to be notified to the user under the control of the control unit 140. The operation unit 139 receives the input operation of the user and transmits the information corresponding to the received input operation to the control unit 140. The display unit 138 may be configured by a touch panel that also serves as an operation unit 139.

For example, the control unit 140 acquires information about the cell culture device 1 from the storage unit 142 based on the identification information 1a read by the reading unit 137. Information about a plurality of cell culture devices 1 having different specifications is stored in advance in the storage unit 142 in a state of being associated with the respective identification information 1a. Further, the information regarding the individual cell culture device 1 is based on the information on the number of the plurality of culture chambers 10 of the cell culture device 1, the information on the position of each of the plurality of culture chambers 10 of the cell culture device 1, and the liquid feed gas 101. Information on the moving pressure of the culture solution between the culture chambers 10 and information on the movement timing of the culture solution between the culture chambers 10 by the liquid feed gas 101 are stored.

For example, the control unit 140 acquires information on the positions of the plurality of culture chambers 10 of the cell culture device 1 from the storage unit 142 based on the identification information 1a. Then, as shown in FIG. 17, the control unit 140 controls the display unit 138 to display the information on the positions of the plurality of culture chambers 10 of the acquired cell culture device 1 so that the user can set it. That is, the control unit 140 controls the display unit 138 to display the information on the position of the standard culture chamber 10 so that the user can change the setting. The user can change the setting of the position information of the standard culture chamber 10 via, for example, the operation unit 139.

In the example of FIG. 17, the control unit 140 controls the display unit 138 to display the information on the position of the culture chamber 10 in the depth direction, that is, the information on the sampling position so that the user can set it. By changing the setting of the sampling position information, for example, sampling of the liquid supernatant in the culture chamber 10 can be performed. In addition, the display unit 138 displays not only the information on the position of the culture chamber 10 in the depth direction but also the information on the horizontal position of the culture chamber 10 (that is, the information on the position in the A direction and the information on the position in the B direction). May be displayed.

The control unit 140 controls to register the position information of the culture chamber 10 acquired based on the identification information 1a as it is in the setting information when the setting is not changed by the user. Further, when the setting is changed by the user, the control unit 140 controls to register the information obtained by adding the setting change by the user to the position information of the culture chamber 10 acquired based on the identification information 1a in the setting information. I do.

Then, at the time of cell culture, the control unit 140 controls the dispensing mechanism 151 and the moving mechanism 152 of the liquid feeding mechanism 150 based on the information on the positions of the plurality of culture chambers 10 of the registered cell culture device 1. .. Specifically, the control unit 140 controls the horizontal movement of the liquid feeding mechanism 150 based on the registered horizontal position information of the culture chamber 10. That is, the control unit 140 horizontally moves the dispensing mechanism 151 by the moving mechanism 152 of the liquid feeding mechanism 150 so that the dispensing mechanism 151 of the liquid feeding mechanism 150 moves to the upper position of the culture chamber 10 to be sucked or discharged. Controls the movement in the direction. Further, the control unit 140 controls the vertical movement of the liquid feeding mechanism 150 based on the registered information on the position of the culture chamber 10 in the depth direction. That is, the control unit 140 determines the stroke amount of the dispensing mechanism 151 of the liquid feeding mechanism 150 based on the information of the position in the depth direction of the culture chamber 10, and sucks or discharges the determined stroke amount. The dispensing mechanism 151 is controlled to be lowered toward the target culture chamber 10 by the moving mechanism 152 of the liquid feeding mechanism 150.

Further, for example, the control unit 140 acquires information on the moving pressure of the culture solution between the culture chambers 10 of the cell culture device 1 by the liquid feed gas 101 from the storage unit 142 based on the identification information 1a. Then, as shown in FIG. 18, the control unit 140 can set the information of the moving pressure of the culture solution between the culture chambers 10 of the cell culture device 1 by the acquired liquid feed gas 101 on the display unit 138. Control the display. That is, the control unit 140 controls the display unit 138 to display the information on the moving pressure of the culture solution between the standard culture chambers 10 so that the user can change the setting. The user can change the setting of the information on the moving pressure of the culture solution between the standard culture chambers 10 via, for example, the operation unit 139.

In the example of FIG. 18, the control unit 140 determines the information on the moving pressure of the culture solution in the culture chamber 10 on the pressurized side as the moving source and the moving pressure of the culture solution between the culture chambers 10 on the non-pressurized side as the moving destination. Information is controlled to be displayed on the display unit 138 so that it can be set by the user. Since the liquid transfer rate between the culture chambers 10 can be changed by changing the setting of the information on the moving pressure of the culture solution, a test in which cells 90 are loaded, such as a test against shear stress, is performed. be able to.

When the setting is not changed by the user, the control unit 140 controls to register the information of the moving pressure of the culture solution between the culture chambers 10 acquired based on the identification information 1a as it is in the setting information. Further, when the setting is changed by the user, the control unit 140 sets the information obtained by adding the setting change by the user to the information on the moving pressure of the culture solution between the culture chambers 10 acquired based on the identification information 1a. Controls registration in information.

Then, at the time of cell culture, the control unit 140 controls the pressure control mechanism 135 based on the information of the moving pressure of the culture solution between the culture chambers 10 of the cell culture device 1 by the registered liquid delivery gas 101. That is, the control unit 140 controls the pressure control mechanism 135 so that the inside of the culture chamber 10 is pressurized by the liquid feed gas 101 according to the moving pressure of the culture solution in the registered culture chamber 10.

Further, for example, the control unit 140 acquires information on the movement timing of the culture solution between the culture chambers 10 of the cell culture device 1 by the liquid feed gas 101 from the storage unit 142 based on the identification information 1a. Then, as shown in FIG. 18, the control unit 140 can set the information of the movement timing of the culture solution between the culture chambers 10 of the cell culture device 1 by the acquired liquid feed gas 101 on the display unit 138. Control the display. That is, the control unit 140 controls the display unit 138 to display the information on the movement timing of the culture solution between the standard culture chambers 10 so that the user can change the setting. The user can change the setting of the information on the movement timing of the culture solution between the standard culture chambers 10 via, for example, the operation unit 139. In the example of FIG. 18, the control unit 140 controls the display unit 138 to display information on the movement time of the culture solution between the culture chambers 10 so that the user can set it.

When the setting is not changed by the user, the control unit 140 controls to register the information of the movement timing of the culture solution between the culture chambers 10 acquired based on the identification information 1a as it is in the setting information. Further, when the setting is changed by the user, the control unit 140 sets the information obtained by adding the setting change by the user to the information on the movement timing of the culture solution between the culture chambers 10 acquired based on the identification information 1a. Controls registration in information.

Then, at the time of cell culture, the control unit 140 controls the pressure control mechanism 135 based on the information on the movement timing of the culture solution between the culture chambers 10 of the cell culture device 1 by the registered liquid delivery gas 101. That is, the control unit 140 pressurizes the inside of the culture chamber 10 with the liquid feed gas 101 so that the culture solution is moved between the culture chambers 10 according to the movement timing of the culture solution in the registered culture chamber 10. Controls the control mechanism 135.

Further, as shown in FIG. 19, the control unit 140 acquires a protocol indicating the processing procedure (culture procedure) of the cell culture device 1 from the storage unit 142 based on the identification information 1a read by the reading unit 137. May be good. In this case, the storage unit 142 stores in advance a protocol indicating the processing procedure (culture procedure) of the cell culture device 1 in a state associated with the identification information 1a. Examples of such a protocol include a standard protocol predetermined for the cell culture device 1 (that is, a culture procedure recommended by the developer of the cell culture device 1), and a protocol registered in advance by the user (that is, the user). The culture procedure that is often performed) can be used.

When the control unit 140 acquires a protocol indicating the processing procedure of the cell culture device 1, the control unit 140 controls the display unit 138 to display the acquired protocol so that the user can set it. That is, the control unit 140 controls the display unit 138 to display the acquired protocol so that the user can change the setting. The user can change the protocol setting via, for example, the operation unit 139.

In the example of FIG. 19, the control unit 140 controls the display unit 138 so that the order and items of the processing procedure according to the protocol can be set by the user. The user can change the order of the presented processing procedures, change, add and delete items, and make detailed settings for each item. As detailed settings for each item, for example, a container, a setting of a reagent placement position, a setting of a reagent addition amount, a setting of a culture solution sampling amount, and the like can be performed. The display unit 138 may display not only one protocol but also a plurality of protocols. In this case, the user can select the protocol to be implemented from a plurality of protocols.

The control unit 140 controls to register the protocol acquired based on the identification information 1a as it is in the setting information when the setting is not changed by the user. Further, when the setting change is made by the user, the control unit 140 controls to register the information obtained by adding the setting change by the user to the protocol acquired based on the identification information 1a in the setting information. Then, at the time of cell culture, the control unit 140 performs control operations such as container transport, reagent adjustment, reagent addition, sampling, and culture solution exchange based on the registered protocol.

As shown in FIG. 20, in the cell culture apparatus 100, the identification information 1b is attached to consumables such as the sample container 161a and the pipette tip 191 and the identification information 1c is attached to the chemicals such as the reagent container 162a. May be good.

In this case, the control unit 140 recognizes the type, number, installation position, and the like of the consumables based on the consumables identification information 1b read by the reading unit 137. Further, the control unit 140 recognizes the type, number, installation position, and the like of the chemicals based on the read identification information 1c of the chemicals. Further, the control unit 140 compares the recognized information on consumables and chemicals with the setting information to detect an error in the installation position or number. When an error in the installation position or number is detected, the control unit 140 warns the user by displaying information on the display unit 138, for example.

(Setting operation of cell culture device)
Next, the setting operation of the cell culture apparatus 100 will be described with reference to FIG. The operation of the cell culture device 100 is controlled by the control unit 140. The setting operation of the cell culture device 100 is an initial setting operation performed before the cell culture experiment of the cell 90 by the cell culture device 1.

As shown in FIG. 21, in step 305, the control unit 140 acquires information based on the

identification information

1a, 1b, and 1c read by the reading unit 137. That is, the control unit 140 acquires information about the cell culture device 1 based on the identification information 1a, acquires information about consumables based on the identification information 1b, and information about chemicals based on the identification information 1c. To get.

In step 306, the control unit 140 causes the display unit 138 to display the setting information. That is, the control unit 140 provides setting information of the position of the culture chamber 10 (see FIG. 17), setting information of movement of the culture solution between the culture chambers 10 (see FIG. 18), protocol setting information (see FIG. 19), and the like. , Displayed on the display unit 138. The user creates setting information of desired culture conditions by using the operation unit 139.

In step 307, the control unit 140 registers the created setting information as the setting information used in the culture experiment and stores it in the storage unit 142. Then, the setting operation of the cell culture device 100 is completed.

(Processing operation of cell culture device)
Next, the processing operation of the cell culture apparatus 100 will be described with reference to FIGS. 22 to 25. The operation of the cell culture device 100 is controlled by the control unit 140.

As shown in FIG. 21, in step 311 the control unit 140 acquires the setting information from the storage unit 142. Based on the device information, consumables information, and liquid information included in the setting information, the control unit 140 sets the moving pressure of the culture solution by the pressure control mechanism 135 and the movement timing of the culture solution by the pressure control mechanism 135 (that is, feeding in circulation culture). Liquid timing) and the position coordinates of the suction position and the discharge position by the dispensing mechanism 151 are acquired. Based on the schedule information (protocol), the control unit 140 acquires the timing of sampling the culture solution, the timing of exchanging the culture solution, and the timing of reagent addition.

In step 312, the control unit 140 controls the liquid feed gas supply unit 120 to prepare the liquid feed gas 101. The liquid feed gas supply unit 120 prepares the liquid feed gas 101 by mixing the air from the air intake unit 121 and the carbon dioxide supplied from the gas source 122. As described above, in the cell culture method of the present embodiment, the step of preparing the liquid feed gas 101 includes the step of mixing the air and the gas to be mixed supplied from the gas source 122.

In step 313, the control unit 140 starts cell culture. The control unit 140 controls the liquid feed gas supply unit 120 and the valve 132 so that the circulation culture is performed according to the setting information. That is, steps 301 to 304 shown in FIG. 2 are carried out. The control unit 140 controls the liquid feed gas supply unit 120 and the valve 132 so that the culture chamber 10 is maintained at the mixed gas concentration and gas pressure (atmospheric pressure) specified in the setting information except during liquid feed. To do. As described above, in the cell culture method of the present embodiment, in the step of moving the liquid in the culture chamber 10 to another culture chamber 10, the liquid is moved by pressurizing the inside of the culture chamber 10 with the supplied liquid feed gas 101. After moving the liquid in the culture chamber 10 to another culture chamber 10, a step of controlling the pressure in the culture chamber 10 to be lower than the pressurized pressure is provided.

Further, the control unit 140 controls each of the first temperature control mechanism 112, the second temperature control mechanism 164, and the third temperature control mechanism 171 so as to maintain each set temperature specified in the setting information. These control actions are continuously executed until the cell culture is stopped in step 321 described later.

In step 314, the control unit 140 determines whether or not it is sampling timing based on the current time and schedule information. If it does not correspond to the sampling timing, the control unit 140 proceeds to step 316. When the sampling timing is met, the control unit 140 controls the sampling operation of the culture solution in step 315, and then proceeds to the process in step 316.

In step 316, the control unit 140 determines whether or not it is the culture solution exchange timing based on the current time and the schedule information. If it does not correspond to the culture solution exchange timing, the control unit 140 proceeds to step 318. When the culture solution exchange timing is applicable, the control unit 140 controls the culture solution exchange operation in step 317, and then proceeds to the process in step 318.

In step 318, the control unit 140 determines whether or not it is the reagent addition timing based on the current time and the schedule information. If it does not correspond to the reagent addition timing, the control unit 140 proceeds to step 320. When the reagent addition timing is applicable, the control unit 140 controls the reagent addition operation in step 319, and then proceeds to the process in step 320.

In step 320, the control unit 140 determines whether or not to end the cell culture. When the control unit 140 ends the cell culture, for example, when the end timing specified in the schedule information is reached, or when the instruction to end the culture is received via the user's operation input using the operation unit 139, etc. to decide. If not, the control unit 140 determines that the cell culture is not finished and returns the process to step 314.

When the control unit 140 determines that the cell culture is finished in step 320, the control unit 140 stops the cell culture in step 321. That is, the control of feeding the culture solution, the control of maintaining the atmosphere of the liquid feed gas 101, and the control of the temperature control, which were started in step 313, are stopped. As a result, the cell culture operation of the cell culture device 100 is completed.

(Sampling operation)
Next, the sampling operation control in step 315 will be described with reference to FIG. 23.

In step 331, the control unit 140 controls the lid opening / closing mechanism 210 to open the lid 33 that covers the culture chamber 10 in which sampling is performed.

In step 332, the control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 to mount the pipette tip 191 and move the pipette tip 191 into the culture chamber 10 for sampling, and move the pipette tip 191 into the inside of the culture chamber 10. Inhale the culture solution. After suction, the control unit 140 retracts the dispensing mechanism 151 from the inside of the culture chamber 10.

In step 333, the control unit 140 controls the lid opening / closing mechanism 210 to close the opened lid 33.

In step 334, the control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 to discharge the culture solution sucked into the pipette tip 191 to the sample container 161a of the container cold insulation unit 160. As described above, the cell culture method of the present embodiment includes a step of feeding the liquid contained in any of the plurality of culture chambers 10 to the sample container 161a. The control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 so that the pipette tip 191 is thrown into the waste port 104 after the excess culture liquid is discharged into the waste liquid port 105 after the culture solution is discharged. ..

(Culture solution exchange operation)
Next, with reference to FIG. 24, control of the exchange operation of the culture solution in step 317 will be described.

In step 341, the control unit 140 controls the container transfer mechanism 180 to transfer the culture solution container 163a stored in the container cold insulation unit 160 to the holding hole 172 of the heating unit 170. The culture solution container 163a is heated by the third temperature control mechanism 171.

In step 342, the control unit 140 controls the lid opening / closing mechanism 210 to open the lid 33 that covers the culture chamber 10 in which the culture solution is exchanged.

In step 343, the control unit 140 controls to discharge the exchanged culture solution. The control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 to mount the pipette tip 191 and move the pipette tip 191 into the culture chamber 10 to suck the culture solution in the culture chamber 10. The control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 so that the sucked culture solution is discharged into the waste liquid port 105 and then the pipette tip 191 is put into the waste port 104.

In step 344, the control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 to mount the pipette tip 191 and suck the new culture solution from the culture solution container 163a warmed by the heating unit 170. ..

In step 345, the control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 to discharge the sucked culture solution into the culture chamber 10. The control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 so that the pipette tip 191 is thrown into the waste port 104 after the excess culture solution is discharged into the waste liquid port 105.

In step 346, the control unit 140 controls the lid opening / closing mechanism 210 to close the opened lid 33.

(Reagent addition operation)
Next, with reference to FIG. 25, control of the reagent addition operation in step 319 will be described.

In step 351 the control unit 140 controls the container transfer mechanism 180 to transfer the reagent container 162a stored in the container cold insulation unit 160 to the holding hole 172 of the heating unit 170. The reagent container 162a is heated by the third temperature control mechanism 171.

In step 352, the control unit 140 controls to suck the reagent in the reagent container 162a. The control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 to mount the pipette tip 191 and suck the reagent from the reagent container 162a warmed by the heating unit 170.

In step 353, the control unit 140 controls the lid opening / closing mechanism 210 to open the lid 33 that covers the culture chamber 10 to which the reagent is added.

In step 354, the control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 to discharge the sucked reagent into the culture chamber 10. As described above, the cell culture method of the present embodiment includes a step of sending the reagent in the reagent container 162a to any of the plurality of culture chambers 10. The control unit 140 controls the moving mechanism 152 and the dispensing mechanism 151 so that the pipette tip 191 is charged into the waste port 104 after the excess reagent is discharged into the waste liquid port 105.

In step 355, the control unit 140 controls the lid opening / closing mechanism 210 to close the opened lid 33.

As described above, in the culture solution exchange and the addition of the reagent, instead of heating the culture solution container 163a and the reagent container 162a with the heating unit 170, a part of the liquid in the culture solution container 163a and the reagent container 162a is sucked. Then, the liquid to be discharged into the culture chamber 10 may be heated by dispensing the liquid into the disposable container 173. For example, when the entire amount of the liquid in the container is not used, or when the reagent is diluted, the liquid is controlled to be dispensed into the disposable container 173.

(Effect of this embodiment)
In this embodiment, the following effects can be obtained.

In the present embodiment, as described above, the culture chamber 10 using the liquid feed gas 101 is based on the reading unit 137 that reads the identification information 1a attached to the cell culture device 1 and the identification information 1a read by the reading unit 137. A control unit 140 that controls the movement of the culture solution between the cells is provided. This saves the trouble of performing the initial setting operation according to the cell culture device 1 used by the user, and controls the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101 suitable for the cell culture device 1 to be used. be able to. As a result, even when cell culture devices 1 having different specifications are used, it is possible to perform control suitable for the cell culture device 1 to be used while eliminating the complexity of initial setting.

Further, in the above embodiment, a further effect can be obtained by configuring as follows.

That is, in the cell culture apparatus 100 of the above embodiment, the control unit 140 sends liquid that is included in the identification information 1a or stored in the storage unit 142 in advance based on the identification information 1a read by the reading unit 137. It is configured to control the acquisition of information regarding the movement of the culture solution between the culture chambers 10 by the gas 101. With this configuration, information on the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101 of the cell culture device 1 to be used can be obtained, so that the liquid feed gas 101 suitable for the cell culture device 1 to be used can be obtained. It is possible to easily control the movement of the culture solution between the culture chambers 10 according to the above.

In the cell culture apparatus 100 of the above embodiment, the control unit 140 uses the liquid feed gas 101 as information regarding the movement of the culture solution between the culture chambers 10a by the liquid feed gas 101 based on the identification information 1a read by the reading unit 137. It is configured to control to acquire at least one of the information on the moving pressure of the culture solution between the culture chambers 10 by 101 and the information on the movement timing of the culture solution between the culture chambers 10 by the liquid feed gas 101. There is. In this way, if the information on the moving pressure of the culture solution between the culture chambers 10 by the liquid feed gas 101 is acquired, the information on the moving pressure of the culture solution between the culture chambers 10 by the acquired liquid feed gas 101 is used for feeding. Initial settings can be made regarding the moving pressure of the culture solution between the culture chambers 10 by the liquid gas 101. As a result, it is possible to reduce the time and effort for the user to perform the initial setting operation regarding the moving pressure of the culture solution between the culture chambers 10 by the liquid feed gas 101. Further, if the information on the movement timing of the culture solution between the culture chambers 10 by the liquid feed gas 101 is acquired, the liquid feed gas is used by using the information on the movement timing of the culture solution between the culture chambers 10 by the acquired liquid feed gas 101. Initial settings can be made regarding the timing of movement of the culture solution between the culture chambers 10 according to 101. As a result, it is possible to reduce the time and effort for the user to perform the initial setting operation regarding the movement timing of the culture solution between the culture chambers 10 by the liquid feed gas 101.

The cell culture device 100 of the above embodiment includes a pressure control mechanism 135 provided in the middle of the gas pipe 130. Further, the control unit 140 has acquired information on the moving pressure of the culture solution between the culture chambers 10 by the liquid feed gas 101 and the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101, which is acquired based on the identification information 1a. It is configured to control the pressure control mechanism 135 based on at least one of the timing information. With this configuration, the user can perform the initial setting operation regarding the moving pressure of the culture solution between the culture chambers 10 by the sending gas 101 and the initial setting regarding the moving timing of the culture solution between the culture chambers 10 by the sending gas 101. The pressure control mechanism 135 can be easily controlled while reducing the labor to be performed.

The cell culture device 100 of the above embodiment includes a display unit 138. Further, the control unit 140 has acquired information on the moving pressure of the culture solution between the culture chambers 10 by the liquid feed gas 101 and the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101, which is acquired based on the identification information 1a. It is configured to control the display unit 138 to display at least one of the timing information so that the user can set it. With this configuration, the information on the moving pressure of the culture solution between the culture chambers 10 by the liquid feed gas 101 and the movement of the culture solution between the culture chambers 10 by the liquid feed gas 101 acquired based on the identification information 1a. At least one of the timing information can reflect the setting change by the user. As a result, cell culture can be performed under desired culture conditions while reducing the time and effort required for the user to perform the initial setting operation.

In the cell culture apparatus 100 of the above embodiment, the control unit 140 controls and cultures the suction material in the culture chamber 10 of the plurality of culture chambers 10 based on the identification information 1a read by the reading unit 137. It is configured to perform at least one of the controls relating to the discharge of the discharged material into the chamber 10. With this configuration, suction in the culture chamber 10 among the plurality of culture chambers 10 suitable for the cell culture device 1 to be used is saved while saving the trouble of performing the initial setting operation according to the cell culture device 1 used by the user. At least one of control regarding suction of the substance and control regarding discharge of the discharged substance into the culture chamber 10 can be performed. As a result, even when cell culture devices 1 having different specifications are used, it is possible to perform control suitable for the cell culture device 1 to be used while eliminating the complexity of initial setting.

In the cell culture apparatus 100 of the above embodiment, the control unit 140 has a plurality of culture chambers included in the identification information 1a or stored in advance in the storage unit 142 based on the identification information 1a read by the reading unit 137. It is configured to control the acquisition of information on each of the ten positions. With this configuration, information on the position of each of the plurality of culture chambers 10 of the cell culture device 1 to be used can be obtained, so that the culture among the plurality of culture chambers 10 suitable for the cell culture device 1 to be used can be obtained. At least one of control regarding suction of the suction material in the chamber 10 and control regarding discharge of the discharge material into the culture chamber can be easily performed.

In the cell culture apparatus 100 of the above embodiment, the dispensing mechanism 151 (first mechanism) that performs at least one of suction of the suctioned material in the culture chamber 10 and discharge of the discharged material into the culture chamber 10 and dispensing. A moving mechanism 152 (second mechanism) for moving the mechanism 151 to the culture chamber 10 to be sucked or discharged is provided. Further, the control unit 140 is configured to control the dispensing mechanism 151 and the moving mechanism 152 based on the information on the positions of the plurality of culture chambers 10 of the cell culture device 1 acquired based on the identification information 1a. ing. With this configuration, the liquid feeding mechanism 150 can be easily controlled while reducing the time and effort for the user to perform the initial setting operation regarding the position of the culture chamber 10 of the cell culture device 1.

The cell culture device 100 of the above embodiment includes a display unit 138. Further, the control unit 140 controls the display unit 138 to display the information on the positions of the plurality of culture chambers 10 of the cell culture device 1 acquired based on the identification information 1a so that the information can be set by the user. It is configured. With this configuration, the setting change by the user can be reflected in the information on the positions of the plurality of culture chambers 10 of the cell culture device 1 acquired based on the identification information 1a. As a result, cell culture can be performed under desired culture conditions while reducing the time and effort required for the user to perform the initial setting operation.

In the cell culture apparatus 100 of the above embodiment, the control unit 140 is configured to control acquisition of a protocol indicating the processing procedure of the cell culture device 1 based on the identification information 1a read by the reading unit 137. There is. With this configuration, the acquired protocol can be used to make initial settings regarding the processing procedure of the cell culture device 1. As a result, it is possible to reduce the time and effort for the user to perform the initial setting operation regarding the processing procedure of the cell culture device 1. This effect is very effective when the processing procedure of the cell culture device 1 is complicated.

The cell culture device 100 of the above embodiment includes a display unit 138. Further, the control unit 140 is configured to control the display unit 138 to display a protocol indicating the processing procedure of the cell culture device 1 acquired based on the identification information 1a so that the user can set it. With this configuration, the setting change by the user can be reflected in the protocol indicating the processing procedure of the cell culture device 1. As a result, cell culture can be performed under desired culture conditions while reducing the time and effort required for the user to perform the initial setting operation.

[Modification example]
It should be noted that the embodiments disclosed this time are exemplary in all respects and are not considered to be restrictive. The scope of the present invention is shown by the scope of claims, not the description of the above-described embodiment, and further includes all modifications (modifications) within the meaning and scope equivalent to the scope of claims.

For example, in the above embodiment, an example of circulating a liquid between a plurality of culture chambers 10 has been shown, but the present invention is not limited to this. In the present invention, instead of circulating the liquid among the plurality of culture chambers 10, it is sufficient to simply move the liquid from one of the culture chambers 10 to another culture chamber 10 connected by the flow path 20.

Further, in the above embodiment, an example in which the culture solution contained in any of the plurality of culture chambers 10 is sent (sampled) to the sample container 161a is shown, but the present invention is not limited to this. In the present invention, sampling is not required. Further, in the above embodiment, an example in which the reagent in the reagent container 162a is sent to any of the plurality of culture chambers 10 (reagent addition) is shown, but the present invention is not limited to this. In the present invention, it is not necessary to add the reagent. Further, in the above embodiment, an example of exchanging the culture solution in the culture chamber 10 has been shown, but the present invention is not limited to this. In the present invention, it is not necessary to replace the culture solution.

Further, when sampling is not performed, the container installation unit 161 may not be provided. When the reagent is not added, the container installation portion 162 may not be provided. When the culture solution is not exchanged, the container installation portion 163 may not be provided. When the container installation units 161 to 163 are not provided, the container cold insulation unit 160, the second temperature control mechanism 164, the third temperature control mechanism 171, the chip rack installation unit 190, and the liquid feeding mechanism 150 may not be provided.

Further, in the above embodiment, an example is shown in which the first temperature control mechanism 112 for controlling the temperature of the cell culture device 1 installed in the installation unit 110 is provided, but the present invention is not limited to this. For example, the temperature of the cell culture device 1 may be adjusted by providing an air conditioner that maintains the internal space of the accommodating portion 102 at a predetermined temperature and adjusting the temperature of the entire internal space of the accommodating portion 102.

Further, in the above embodiment, an example in which the leak pipe 131 connected to the cell culture device 1 and discharging the gas in the culture chamber 10 is provided is shown, but the present invention is not limited to this. The leak pipe 131 may be connected to the gas pipe 130.

Further, in the above embodiment, an example in which the connector mechanism 113 is provided is shown, but the present invention is not limited to this. In the present invention, the connector mechanism 113 may not be provided. For example, when the cell culture device 1 is installed in the installation unit 110, the user may perform the work of connecting the gas pipes 130 one by one to the gas supply port 31. The same applies to the discharge mechanism 116, and the discharge mechanism 116 does not have to be provided. The user may perform the work of connecting the leak pipes 131 to the gas discharge port 36 one by one.

Further, in the above embodiment, information on the position of each of the plurality of culture chambers 10 of the cell culture device 1, information on the moving pressure of the culture solution between the culture chambers 10 of the cell culture device 1 by the liquid feed gas 101, or feeding. An example has been shown in which information on the movement timing of the culture solution between the culture chambers 10 of the cell culture device 1 by the liquid gas 101 can be set by the user and displayed on the display unit 138, but the present invention is not limited to this. In the present invention, information on the position of each of the plurality of culture chambers 10 of the cell culture device 1, information on the moving pressure of the culture solution between the culture chambers 10 of the cell culture device 1 by the liquid feed gas 101, or the liquid feed gas 101. Information on the movement timing of the culture solution between the culture chambers 10 of the cell culture device 1 according to the above can be set by the user, and it is not necessary to display it on the display unit 138. For example, information on the position of each of the plurality of culture chambers 10 of the cell culture device 1, information on the moving pressure of the culture solution in the culture chamber 10 of the cell culture device 1 by the liquid feed gas 101, or cell culture by the liquid feed gas 101. The information on the movement timing of the culture solution between the culture chambers 10 of the device 1 may be used as it is as the setting information.

Further, in the above embodiment, an example in which the display unit 138 is provided is shown, but the present invention is not limited to this. In the present invention, it is not necessary to provide the display unit 138. For example, the information may be displayed on an external display device.

[Aspect]
It will be understood by those skilled in the art that the above exemplary embodiments are specific examples of the following embodiments.

(Item 1)
An installation unit in which a cell culture device having a plurality of culture chambers including a first culture chamber and a second culture chamber is installed, and
A liquid feed gas supply unit for supplying a liquid feed gas for moving the culture solution between the first culture chamber and the second culture chamber, and a liquid feed gas supply unit.
A gas pipe that detachably connects the cell culture device installed in the installation unit and the liquid feed gas supply unit, and
A reading unit that reads the identification information attached to the cell culture device,
A cell culture apparatus including a control unit that controls the movement of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas based on the identification information read by the reading unit. ..

(Item 2)
Based on the identification information read by the reading unit, the control unit has the first culture chamber and the second culture using the liquid feed gas, which is included in the identification information or stored in the storage unit in advance. The cell culture apparatus according to item 1, wherein the cell culture apparatus is configured to control the acquisition of information regarding the movement of the culture medium between chambers.

(Item 3)
Based on the identification information read by the reading unit, the control unit uses the liquid feed gas as information regarding the movement of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas. Of the information on the moving pressure of the culture solution between the first culture chamber and the second culture chamber and the information on the movement timing of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas. The cell culture apparatus according to item 2, wherein the cell culture apparatus is configured to control the acquisition of at least one of the above.

(Item 4)
Further provided with a pressure control mechanism provided in the middle of the gas pipe,
The control unit obtains information on the moving pressure of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas, and the first one by the liquid feed gas, which is acquired based on the identification information. The cell culture apparatus according to item 3, wherein the pressure control mechanism is controlled based on at least one of information on the movement timing of the culture solution between the culture chamber and the second culture chamber.

(Item 5)
With an additional display
The control unit obtains information on the moving pressure of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas, and the first one by the liquid feed gas, which is acquired based on the identification information. Item 3 or 4 is configured to control display on the display unit so that at least one of the information on the movement timing of the culture solution between the culture chamber and the second culture chamber can be set by the user. The cell culture apparatus according to.

(Item 6)
The control unit controls the suction of the suction material in the culture chamber among the plurality of culture chambers and the control of the discharge material into the culture chamber based on the identification information read by the reading unit. The cell culture apparatus according to any one of items 1 to 5, which is configured to perform at least one of the above.

(Item 7)
The control unit acquires information on the position of each of the plurality of culture chambers included in the identification information or stored in advance in the storage unit based on the identification information read by the reading unit. The cell culture apparatus according to item 6, wherein the cell culture apparatus is configured to perform the above.

(Item 8)
A first mechanism that sucks the suctioned material in the culture chamber and discharges the discharged material into the culture chamber, and a second mechanism that moves the first mechanism to the culture chamber to be sucked or discharged. With a mechanism,
The control unit is configured to control the first mechanism and the second mechanism based on the information on the positions of the plurality of culture chambers of the cell culture device acquired based on the identification information. The cell culture apparatus according to item 7.

(Item 9)
With an additional display
The control unit is configured to control the display unit to display information on the position of each of the plurality of culture chambers of the cell culture device acquired based on the identification information so that the user can set it. The cell culture apparatus according to item 7 or 8.

(Item 10)
The control unit is configured to control acquisition of a protocol indicating a processing procedure of the cell culture device based on the identification information read by the reading unit, any one of items 1 to 9. The cell culture apparatus according to the section.

(Item 11)
With an additional display
The control unit is configured to control the display on the display unit so that the protocol indicating the processing procedure of the cell culture device acquired based on the identification information can be set by the user. The cell culture apparatus described.

1

Cell culture device

10, 10a, 10b, 10c, 10d culture chamber (first culture chamber, second culture chamber)
100 Cell culture device 101 Liquid supply gas 110 Installation unit 120 Liquid supply gas supply unit 130 Gas piping 135 Pressure control mechanism 137 Reading unit 138 Display unit 140 Control unit 150 Liquid supply mechanism 151 Dispensing mechanism (1st mechanism)
152 Moving mechanism (second mechanism)

Claims

An installation unit in which a cell culture device having a plurality of culture chambers including a first culture chamber and a second culture chamber is installed, and
A liquid feed gas supply unit for supplying a liquid feed gas for moving the culture solution between the first culture chamber and the second culture chamber, and a liquid feed gas supply unit.
A gas pipe that detachably connects the cell culture device installed in the installation unit and the liquid feed gas supply unit, and
A reading unit that reads the identification information attached to the cell culture device,
A cell culture apparatus including a control unit that controls the movement of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas based on the identification information read by the reading unit. ..
Based on the identification information read by the reading unit, the control unit has the first culture chamber and the second culture using the liquid feed gas, which is included in the identification information or stored in the storage unit in advance. The cell culture apparatus according to claim 1, wherein the cell culture apparatus is configured to control the acquisition of information regarding the movement of the culture medium between chambers.
Based on the identification information read by the reading unit, the control unit uses the liquid feed gas as information regarding the movement of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas. Of the information on the moving pressure of the culture solution between the first culture chamber and the second culture chamber and the information on the movement timing of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas. The cell culture apparatus according to claim 2, wherein the cell culture apparatus is configured to control the acquisition of at least one of the above.
Further provided with a pressure control mechanism provided in the middle of the gas pipe,
The control unit obtains information on the moving pressure of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas, and the first one by the liquid feed gas, which is acquired based on the identification information. The cell culture apparatus according to claim 3, wherein the pressure control mechanism is controlled based on at least one of information on the movement timing of the culture solution between the culture chamber and the second culture chamber.
With an additional display
The control unit obtains information on the moving pressure of the culture solution between the first culture chamber and the second culture chamber by the liquid feed gas, and the first one by the liquid feed gas, which is acquired based on the identification information. The third aspect of claim 3 is configured to control display of at least one of the information on the movement timing of the culture solution between the culture chamber and the second culture chamber on the display unit so as to be set by the user. The cell culture apparatus according to the above.
The control unit controls the suction of the suction material in the culture chamber among the plurality of culture chambers and the control of the discharge material into the culture chamber based on the identification information read by the reading unit. The cell culture apparatus according to claim 1, wherein the cell culture apparatus is configured to perform at least one of the two.
The control unit acquires information on the position of each of the plurality of culture chambers included in the identification information or stored in advance in the storage unit based on the identification information read by the reading unit. The cell culture apparatus according to claim 6, wherein the cell culture apparatus is configured to perform the above.
A first mechanism that sucks the suctioned material in the culture chamber and discharges the discharged material into the culture chamber, and a second mechanism that moves the first mechanism to the culture chamber to be sucked or discharged. With a mechanism,
The control unit is configured to control the first mechanism and the second mechanism based on the information on the positions of the plurality of culture chambers of the cell culture device acquired based on the identification information. The cell culture apparatus according to claim 7.
With an additional display
The control unit is configured to control the display unit to display information on the position of each of the plurality of culture chambers of the cell culture device acquired based on the identification information so that the user can set it. The cell culture apparatus according to claim 7.
The cell culture according to claim 1, wherein the control unit controls to acquire a protocol indicating a processing procedure of the cell culture device based on the identification information read by the reading unit. apparatus.
With an additional display
10. The control unit is configured to control display on the display unit so that a protocol indicating a processing procedure of the cell culture device acquired based on the identification information can be set by the user. The cell culture apparatus according to.