Cell culture system

Abstract

The present application relates to a cell culture system, and relates to an automatic culture device using a single-use closed system flow path for culturing cells or tissues, and to realize reduction of manufacturing cost and high concentration of the device. The cell culture system is provided with an automatic culture device provided with a plurality of closed system flow paths and a plurality of culture devices which can be set and detached, and an information processing device provided with: an input device which accepts at least one data selected from a group consisting of data of an identifier of a patient, data related to a transplantation method, data related to a cell type, data related to a required cell number, and data related to a treatment plan as an input; a calculation device which selects a cell culture method, the culture device, and the closed system flow path to be used from options related to the cell culture method, the culture device, and the closed system flow path based on the input data; and an output device which outputs a number of the closed system flow path to be used and a number of the culture device to be used.

Description

TECHNICAL FIELD

[0001] The present application relates to a cell culture system. BACKGROUND

[0002] As a root therapy for a disease for which no therapy has been available in the past, regenerative medicine using a regenerative tissue or the like manufactured using cells as a material to restore the function of an organ or the like is expected. For various therapeutic target tissues such as skin, cornea, esophagus, heart, bone, and cartilage, the number of clinical application cases is rapidly increasing. In a manufacturing process of a regenerative tissue, stem cells are isolated from a biological sample collected from a patient himself or another person, and the isolated stem cells are proliferated or organized. This process is performed in a cell processing facility (CPF) in accordance with a standard instruction manual (SOP) of a proper manufacturing guideline (GMP) that satisfies a guideline for manufacturing management and quality management of pharmaceutical products and the like.

[0003] The use of the CPF requires a large amount of cost and personnel with specialized culture techniques. In addition, the manufacturing process of a regenerative tissue is centered on manual work, and thus there is a limit to the amount of manufactured regenerative tissues. Low productivity and high manufacturing cost are obstacles to the popularization of regenerative medicine, and automation of culture work that requires labor and cost in the manufacturing process is particularly expected, and thus labor-saving, cost reduction, and mass production of regenerative tissues can be achieved.

[0004] In addition, among products that have been marketed and products under development, regenerative tissues for autologous transplantation are more than those for allogenic transplantation. Autologous transplantation using a patient's own cells is expected to have a good therapeutic result because of a low possibility of rejection, and is considered to be in high demand from the viewpoint of improving the QOL of patients. In autologous transplantation, a regenerative tissue is manufactured for each patient, and thus a variety of regenerative tissues can be produced in small amounts. At this time, if manufacturing using an automated culture device is assumed, manufacturing of one patient's amount in one automated culture device is preferred in order to avoid the risk of biological contamination. On the other hand, if manufacturing of cells of a plurality of patients is assumed, small size and high concentration are also important. Therefore, low manufacturing cost and high concentration of an automated culture device for autologous transplantation are important.

[0005] There are two kinds of automatic culture devices, an open system automatic culture device and a closed system automatic culture device. In the open system automatic culture device, open system culture containers such as culture containers of a type in which a lid is opened and closed, culture plates, and the like, which are usually used in manual culture, are automatically handled in a closed space provided with a multi-joint robot or the like. The closed space can be decontaminated with a decontamination gas or the like. On the other hand, the closed system automatic culture device automatically handles a closed system flow path having a closed space. The closed system flow path is a state in which closed system culture containers are always connected through a flow path tube or the like, and the inside is sterilized in advance by gamma-ray sterilization or the like.

[0006] In the case of comparing the open system automatic culture device and the closed system automatic culture device, in the open system automatic culture device, the open system culture containers are taken out of the open system automatic culture device after the production of the regenerative tissue and are transferred to the next process, but in the transfer of the open system culture containers, there is a risk of biological contamination caused by leakage of the culture medium or the like.

[0007] In the closed system automatic culture device, the closed system culture containers are taken out of the closed system automatic culture device after the production of the regenerative tissue and are transferred to the next process, but in the transfer of the closed system culture containers, there is no leakage of the culture medium, and thus the risk of biological contamination is lower than in the open system automatic culture device using the open system culture containers. In addition, since the open system automatic culture device is provided with a multi-joint robot or the like in a closed space, the device is mostly large. Therefore, in the case where a plurality of open system automatic culture devices are used in a state of being gathered, a room in which the CPF of the device is housed needs to be a large-scale room. The closed system automatic culture device makes only the inside of the closed system flow path a sterile state, and thus the device can be downsized. Therefore, the closed system automatic culture device is easier to use than the open system automatic culture device in a state in which a plurality of closed system automatic culture devices are gathered.

[0008] As an example of the closed system automatic culture device, a device having one layer of closed system culture containers and always connected with a flow path for supplying or discharging a culture medium or the like is disclosed (see Patent Document 1). In the case of sending a culture medium or the like, a valve provided outside the closed system flow path is adjusted, and a pump is operated. The closed system flow path is designed to be single-use in order to avoid biological contamination. In addition, it is also possible to use in a state in which a plurality of devices are gathered. In addition, a closed system automatic culture device provided with a plurality of flow paths, branches, and the like in the closed system culture containers is also disclosed (see Patent Document 2). In this device, by selecting an appropriate flow path or branch from among the plurality of flow paths and branches, an automatic culture protocol corresponding to the selected flow path or branch can be implemented.

[0009] Patent Document 1: Japanese Patent Application Publication No. 2007-312668

[0010] Patent Literature 2: US2019 / 0330584 SUMMARY

[0011] An object of the present application is to provide a new cell culture system.

[0012] One embodiment of the present application is a cell culture system including an automatic culture device and an information processing device, wherein the automatic culture device includes a plurality of closed system flow paths and a plurality of culture devices, the information processing device includes an input device that accepts at least one data selected from a group consisting of data of an identifier of a patient, data related to a transplantation method, data related to a cell type, data related to a number of cells required, and data related to a treatment plan as an input, an arithmetic device that selects a cell culture method, the culture device, and the closed system flow path to be used from options related to the cell culture method, the culture device, and the closed system flow path based on the data input, and an output device that outputs a number of the closed system flow path to be used and a number of the culture device to be used. The closed system flow path can include a valve for opening and closing a flow path tube, a pump for sending a liquid or a gas in the flow path tube, a culture medium container for holding fresh culture medium, and a culture supernatant container for holding culture supernatant after culture. One closed system flow path can be provided for a plurality of culture devices. The output device can output a work order procedure related to the closed system flow path to be used. The output device can output a work order procedure related to the culture device selected.

[0013] According to the automatic culture device of the present application, a plurality of culture devices use a closed system flow path having high versatility and a culture program using a plurality of culture devices, thereby improving the concentration of the automatic culture device, and as a result, the manufacturing cost can be reduced. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 FIG. 1 is a diagram showing the structure of a culture device according to one embodiment of the present application.

[0015] Figure 2 FIG. 2 is a diagram showing a state in which a closed system flow path is provided in the culture device according to one embodiment of the present application.

[0016] Figure 3 FIG. 3 is a diagram showing four closed system flow paths provided in the culture device according to one embodiment of the present application.

[0017] Figure 4 FIG. 4 is a diagram showing the structure in which a plurality of culture devices are stacked according to one embodiment of the present application.

[0018] Figure 5 is a view showing a state in which a closed system flow path is provided to each culture device in a structure in which a plurality of culture devices are stacked in one embodiment of the present application.

[0019] Figure 6 is a view showing a state in which one closed system flow path is provided to every two or three culture devices in a structure in which a plurality of closed system automatic culture devices are stacked in one embodiment of the present application.

[0020] Figure 7-1 is a view showing a closed system flow path provided to a plurality of culture devices in one embodiment of the present application.

[0021] Figure 7-2 is a view showing a closed system flow path provided to a plurality of culture devices in one embodiment of the present application.

[0022] Figure 8 is a view showing culture information related to a culture device and a closed system flow path used in culture in one embodiment of the present application.

[0023] Figure 9 is a view showing flow of culture information related to a culture device and a closed system flow path used in culture in one embodiment of the present application.

[0024] Figure 10 is a view showing a screen example of a culture program in one embodiment of the present application.

[0025] Figure 11 is a view showing a control mechanism of an automatic culture device in one embodiment of the present application.

[0026] Figure 12 is a view showing a flow when a closed system automatic culture device operates in one embodiment of the present application.

[0027] Figure 13 is a view showing a structure of a culture device in which a culture medium container is provided to a device side surface at room temperature without a refrigeration chamber in another embodiment of the present application.

[0028] Figure 14 is a view showing a structure of a culture device in which a culture medium container is provided to a device side surface at room temperature without a refrigeration chamber and a flow path mechanism portion in another embodiment of the present application. DETAILED DESCRIPTION

[0029] The following describes preferred embodiments of the present application using the drawings, but the application is not necessarily limited thereto. Furthermore, the objects, features, advantages and concepts of the present application will be apparent from the description of the application, and those skilled in the art can easily reproduce the present application based on the description. The embodiments and specific examples of the application described below represent preferred embodiments of the present application, and are shown for illustration or explanation, and do not limit the present application thereto. Various changes and modifications can be made based on the description of the present application, and it will be apparent to those skilled in the art that such changes and modifications are within the intent and scope of the present application.

[0030] The cell culture system of the present application includes an information processing device and an automatic culture device.

[0031] The automatic culture device includes a plurality of closed system flow paths and a plurality of culture devices that can be attached and detached. The information processing device includes an input device that accepts at least one data selected from a group consisting of data of an identifier of a patient, data related to a transplantation method, data related to a cell type, data related to a number of cells required, and data related to a treatment plan as an input, a calculation device that determines a cell culture method, a culture device, and a closed system flow path to be used based on the input data from options related to the cell culture method, the culture device, and the closed system flow path, and an output device that outputs a number of the closed system flow path to be used and a number of the culture device to be used.

[0032] The closed system flow path used in the culture includes a closed system culture container, a culture medium container, a culture supernatant container, and the like. Various closed system flow paths that differ in the type, number, and the like of the closed system culture container and the like can be used according to the cell culture method corresponding to the cell type to be cultured, the cell type after culture, and the form of the regenerated tissue, and the like. That is, in one automatic culture device, culture using a plurality of closed system flow paths based on different cell culture methods can be performed.

[0033] The information processing device includes an input device, a calculation device, and an output device. In order to use a plurality of closed system flow paths for a plurality of culture devices and perform a cell culture method according to each closed system flow path, a program using the plurality of culture devices is used in the information processing device. The program has an electronic tag that can associate each culture device and each closed system flow path with a cell culture method. Based on at least one data selected from a group consisting of data of an identifier of a patient, data related to a transplantation method, data related to a cell type, data related to a number of cells required, and data related to a treatment plan received by the input device, the calculation device determines a cell culture method, a culture device, and a closed system flow path to be used. The output device outputs a job order sheet corresponding to the cell culture method to be used.

[0034] Thus, by automatically selecting a flow path for culturing from among a plurality of culture devices and a plurality of closed system flow paths, culturing of cells is performed, which enables an increase in the concentration and a reduction in the manufacturing cost of the cell culturing system.

[0035] Hereinafter, with reference to the drawings, an embodiment of a cell culturing system that automatically performs culturing using a closed system flow path will be described, but the cell culturing system of the present application is not limited to the following embodiment, and various modifications can be made by those skilled in the art.

[0036] <Embodiment I>

[0037] In the present Embodiment I, a cell culturing system that automatically performs culturing using a closed system flow path will be described. Figure 1 , Figure 2

[0038] Each culture device is provided with a culture chamber 101 for culturing cells at 37°C, which is the culture temperature, a flow path mechanism section 102 having a liquid feeding function, and a refrigeration section 103 Figure 1 ). The flow path mechanism section 102 is provided with a pump 104 and a solenoid valve 105. The refrigeration section 103 stores a culture medium container having a culture medium stored therein and a culture supernatant container for recovering culture supernatant. As to the shape of these containers, a bottle or a bag can be exemplified.

[0039] Figure 2 represents a flow path mechanism section 102 of a culture device 100, and a closed system flow path 200 is connected to the flow path mechanism section 102. Figure 1 ​The inside of the culture device is provided with a closed system flow path. The closed system culture container 201 in which cells are cultured is provided inside the incubator 101. The number of closed system culture containers 201 can be one or a plurality. The closed system culture container 201 is always connected to the culture medium container 202, culture supernatant container, and the like located in the refrigeration section 103 via the flow path tube 203. The flow path tube 203 is installed in the incubator 101, and therefore a notch section 106, which is a space through which the flow path tube 203 passes, is provided in the incubator 101. The notch section 106 can be provided in the door of the incubator 101 or in the main body. The notch section 106 has airtightness, and therefore the inside of the incubator 101 can be maintained at an appropriate temperature, CO2 concentration, and humidity in a state in which the flow path tube 203 is provided. For example, the flow path tube 203 is made of a plastic material, and can be embedded in the gap of the notch section 106 in a pressed manner. Alternatively, in the notch section 106 in which the flow path tube 203 passes, the gap can be embedded with a jig having plasticity. Furthermore, in a state in which the flow path tube 203 is not provided, the gap of the notch section 106 can be embedded with a jig having plasticity and embedded in the gap of the notch section 106. Thus, when the inside of the incubator 101 is decontaminated with a decontaminating gas such as hydrogen peroxide, the inside of the incubator 101 can be sealed even if the flow path tube 203 is not provided.

[0040] Next, the use of the closed system flow path will be described. Figure 3 Embodiments of the closed system flow path will be described. In the present embodiment, four types of closed system flow paths can be used in one automatic culture device, but the types and number of closed system flow paths are not limited thereto, and a closed system flow path in which a part of the structure is changed can also be used. Depending on the type of the closed system flow path, the cell culture method that can be performed is different, and the culture protocol for culture is also different.

[0041] Figure 3(A) of FIG. 1 is a flow path that enables adherent culture of stem cells such as iPS cells, mesenchymal stem cells, and the like. In this configuration, both discharge of culture supernatant used in culture and supply of new culture medium can be performed in medium exchange. By adjusting the height of the tube on the discharge side within the closed system culture vessel 301, medium exchange at an arbitrary ratio can also be performed. The filter-attached closed system culture vessel 301 is provided within the incubator 101. The filter-attached closed system culture vessel 301 has a filter 302 for gas exchange between the gas phase within the incubator 101 and the gas phase within the filter-attached closed system culture vessel 301. A culture medium container 306 containing fresh culture medium, a culture supernatant container 307 are provided within the refrigeration section 103. All constituent elements are always connected by the flow path tube 303. The flow path tube 303 is given a driving force for transporting culture medium and the like from the outside by a pump 304, and the liquid feeding direction at the time of liquid feeding is selected by a solenoid valve 305, whereby liquid feeding of culture medium and the like is performed. The solenoid valve 305 is closed during culture. Although the filter-attached closed system culture vessel 301 is used, a closed system culture vessel 308 without a filter can also be used. Figure 3 (B) of FIG. 1 is a closed system culture vessel 308 with a gas-permeable membrane used in (A). As a culture protocol, only supply of new culture medium can be performed, and culture can be performed without using the culture supernatant container 307.

[0042] As an example, Figure 3 (B) of FIG. 1 is a flow path that enables suspension culture of T cells. As a culture protocol, only supply of culture medium is performed. For the two closed system culture vessels 308, there is a common culture medium container 306. The closed system culture vessel 308 with a gas-permeable membrane is provided within the incubator 101. The closed system culture vessel 308 with a gas-permeable membrane has a gas-permeable membrane 309 for gas exchange between the gas phase within the incubator 101 and the gas phase within the closed system culture vessel 308 with a gas-permeable membrane. A culture medium container 306 containing fresh culture medium is provided within the refrigeration section 103. At the time of liquid feeding, the two solenoid valves 305 determine the liquid feeding direction. The solenoid valve 305 for the closed system culture vessel 308 with a gas-permeable membrane that is the liquid feeding target is opened, and the solenoid valve 305 for the closed system culture vessel 308 with a gas-permeable membrane that is not the liquid feeding target is closed. The solenoid valve 305 is closed during culture. Furthermore, in the case of using the closed system culture vessel 308 with a gas-permeable membrane, a closed system culture vessel 308 without a gas-permeable membrane can also be used. Figure 3 (B) of FIG. 1, the closed system culture vessel 308 with a gas-permeable membrane has a gas-permeable membrane 309 at two positions of the lid and the lower portion, but can also be single-sided. Although the closed system culture vessel 308 with a gas-permeable membrane is used, a closed system culture vessel 308 without a gas-permeable membrane can also be used. Figure 3 (A) of FIG. 1 is a filter-attached closed system culture vessel 301 used in (A).

[0043] As an example, Figure 3(C) is a flow path capable of suspension-adherent culture of T cells. As the culture protocol, only the supply of culture medium is performed. For the 2 closed-system culture containers 308, there are culture medium containers 306 each filled with fresh culture medium. That is, 2 closed-system flow paths are used. Furthermore, in the present figure, the closed-system culture container 308 with a gas-permeable membrane has a gas-permeable membrane 309 at 2 positions of the lid and the lower portion, but can also be single-sided. Although the closed-system culture container 308 with a gas-permeable membrane is used, a closed-system culture container without a gas-permeable membrane can also be used Figure 3 The filter external closed-system culture container 301 used in (A).

[0044] As an example, Figure 3 (D) is a flow path capable of adherent culture of stem cells such as iPS cells and mesenchymal stem cells, and suspension culture of T cells. As the culture protocol, perfusion culture is performed while maintaining the liquid volume constant, with the old culture medium being discharged while the new culture medium is supplied. The perfusion-culture closed-system culture container 310 is provided in the incubator 101. The perfusion-culture closed-system culture container 310 has a gas-permeable membrane 309 for gas exchange between the gas phase inside the incubator 101 and the gas phase inside the perfusion-culture closed-system culture container 310. The culture medium container 306 filled with fresh culture medium, and the culture supernatant container 307 are provided inside the refrigeration section 103. Furthermore, in the present figure, the closed-system culture container 308 with a gas-permeable membrane has a gas-permeable membrane 309 at 1 position of the upper portion, but can be provided at the lower portion or both.

[0045] Next, the case where the culture device is used in a stacked state will be described. Figure 4 , Figure 5 , Figure 6 The case where the culture device is used in a stacked state will be described.

[0046] Figure 4 In (E), as examples, a state where 2 culture devices are stacked and a state where 3 culture devices are stacked are shown.

[0047] The number of stacked devices can be selected by the user according to the purpose or the like. Figure 5 is a figure of culture of 1 culture device based on 1 closed-system flow path. Figure 6 (A) is a figure of culture using 2 culture devices based on 1 closed-system flow path. Figure 6 (B) is a figure of culture using 3 culture devices based on 1 closed-system flow path. Although not shown here, culture using 4 or more culture devices based on 1 closed-system flow path is also possible. In this way, in the automatic culture device of the present disclosure, a plurality of culture devices can be connected by 1 closed-system flow path and cultured. As a result, culture can be performed on an arbitrary number of culture containers, and cells can be mass-produced.

[0048] The closed system flow path has a closed system culture container 201, a culture medium container, and a culture supernatant container corresponding to the number of culture devices. By opening and closing the electromagnetic valve 105, the liquid feeding direction is switched to the culture container that becomes a liquid feeding target, and then the pump 104 is operated to feed the liquid to each culture container. The culture medium container is provided in each refrigeration chamber, and by opening and closing the electromagnetic valve 105, the liquid feeding direction can be switched to the culture medium container that becomes a liquid feeding target, and then the pump 104 is operated to feed the liquid from the culture medium container. In the case of long-term culture, the culture medium has a use-by date, and therefore a new culture medium can be provided using a sterile connection portion. Since the amount of culture medium required for culture is large, the culture medium can also be provided separately in each refrigeration chamber.

[0049] Next, using FIG. 7, another embodiment of the automatic culture device used in a state in which culture devices are stacked will be described. FIG. 7 shows an automatic culture device used in a state in which two devices are stacked and a state in which three devices are stacked. Depending on the type of closed system flow path, the culture protocol that can be implemented also differs. Therefore, the user can select the closed system flow path, and the culture method is selected in accordance with the selected closed system flow path.

[0050] As an example, (A) and (B) of FIG. 7 are automatic culture devices for culturing stem cells such as iPS cells and mesenchymal stem cells. (A) of FIG. 7 is an automatic culture device used in a state in which two culture devices are stacked. (B) of FIG. 7 is an automatic culture device used in a state in which three culture devices are stacked. As the culture protocol, either the discharge of the culture supernatant for culture or the supply of new culture medium can be performed in the culture medium exchange. By adjusting the height of the tube on the discharge side inside the closed system culture container 301, culture medium exchange at an arbitrary ratio can also be performed. The filter-external closed system culture container 301 is provided in the incubator 101. The culture medium container 306 is provided inside the refrigeration section 103. Furthermore, in (A) of FIG. 7, two culture medium containers 306 and two culture supernatant containers are provided, but the number of culture medium containers 306 can be one or three depending on the required amount of culture medium.

[0051] The same applies to the culture supernatant container. Although the filter-attached closed system culture container 301 is used, the closed system culture container 308 with a gas-permeable membrane can also be used. As the culture protocol, only the supply of fresh medium can be performed to culture without using the culture supernatant container. The same applies to the automatic culture device of (B) of FIG. 7. As examples, (C) and (D) of FIG. 7 are flow paths for suspension culture of T cells. (C) of FIG. 7 is an automatic culture device used in a state where two culture devices are stacked. (D) of FIG. 7 is an automatic culture device used in a state where three culture devices are stacked. As the culture protocol, only the supply of medium is performed. The closed system culture container 308 with a gas-permeable membrane is provided in the incubator 101. The medium container 306 is provided in the refrigeration section 103. In addition, two medium containers 306 are provided in (C) of FIG. 7, but the number of medium containers 306 can be one or three depending on the required amount of medium. Although the closed system culture container 308 with a gas-permeable membrane is used, the filter-attached closed system culture container 301 can also be used. The same applies to the automatic culture device of (D) of FIG. 7.

[0052] In the case of culturing cells of a plurality of patients, it is preferable to provide the culture devices in a state of aggregation in a cell conditioning room in one room. In each culture device, cells of different patients can be cultured by respective culture protocols. The culture protocols differ depending on the type of cells to be cultured, and the schedule of the culture protocols differs depending on the schedule of transplantation and the like treatment which differs for each patient. In each closed system flow path used, the type and number of culture containers differ depending on the type of cells to be cultured and the mode of transplantation. Therefore, it is necessary to appropriately select the protocol, closed system flow path, and culture device to be used for each patient from among a plurality of culture protocols, closed system flow paths, and culture devices.

[0053] According to Figure 8 The culture information concerning a plurality of culture devices and a plurality of closed system flow paths used in culture possessed by the information processing device is described. Figure 8 The table shows the generation and acquisition of culture information generated in each process. In addition, the flow of culture information concerning a plurality of culture devices and a plurality of closed system flow paths used in culture, and the relationship between the culture protocol for processing the culture information and the culture information are shown in Figure 9 The flow of culture information concerning a plurality of culture devices and a plurality of closed system flow paths used in culture, and the relationship between the culture protocol for processing the culture information and the culture information are shown in

[0054] Starting from the patient's visit to the hospital, as the treatment method is decided, information relating to the cells to be manufactured is generated. The output device outputs and the display device displays the work order procedure relating to the extraction of the closed system flow path to be used, the determination of the selected culture device. When each item is decided and input to the input device, the operation device limits the options in the subsequent selection according to the decided content, the output device outputs the options to the display device, and the display device displays the options. For example, when the type of cells to be manufactured is decided and input to the input device, the operation device limits the type of culture containers that can be used for culture to a type corresponding to the type of cells, the output device outputs the options to the display device, and the display device displays the options. In addition, when the type, number, and transplantation method of the cells to be manufactured are decided and input to the input device, the operation device limits the manufacturing method according to the conditions.

[0055] In the decision of the manufacturing method, first, the type of closed system flow path to be used is decided and input to the input device. This depends on the type and number of culture containers to be used. Next, the culture device to be used is decided and input to the input device. In addition, the culture period differs depending on the cells to be manufactured, that is, the operation period of the automatic culture device also differs. In addition, the culture schedule varies for each patient. Improving the operation rate of the device and manufacturing as many cells as possible for each patient reduces the manufacturing cost, so it is also possible to select a culture device used in a manner in which the operation rate is as high as possible. When the closed system flow path to be used is moved out from the warehouse, the batch number of the moved-out closed system flow path is displayed on the work order procedure on the display device. An electronic tag or 2-dimensional barcode having the batch number information is installed in advance in the closed system flow path, and it is confirmed that the batch number displayed in the work procedure book coincides with the information of the electronic tag or 2-dimensional barcode of the selected closed system flow path. When the closed system flow path to be used is set in the culture device, the device number of the culture device to be used is displayed on the work order procedure on the display device. An electronic tag or 2-dimensional barcode having the device number information is installed in advance in the culture device, and it is confirmed that the batch number displayed in the work procedure book coincides with the information of the electronic tag or 2-dimensional barcode of the selected culture device. By the above method, in the selection of the closed system flow path and the culture device to be used, selection errors are avoided. After manufacturing, the quality evaluation results are used to determine whether or not the manufactured cells are used for treatment.

[0056] Figure 10 An example of a display screen indicating a work order procedure driven by a culture program. Figure 10(A) is a screen for selecting a cell type in selection relating to a treatment method. Before the selection of the cell type, in this example, a cancer immunotherapy is selected as the treatment method, and therefore, in the selection of the cell type, only the cell types that can be used in the cancer immunotherapy are displayed as options from which to select. Cell types that are not used in the cancer immunotherapy are not displayed, thereby avoiding errors in selection. Figure 10 (B) is a screen for selecting a type of culture vessel in selection relating to a culture method. Before the selection of the type of culture vessel, since a suspension cell is selected as the culture method, in the selection of the type of culture vessel, only the types of culture vessels that can be used in the suspension cell are displayed as options from which to select. Figure 10 (C) is also the same selection form. Figure 10 (D) is a screen that displays all the selected information.

[0057] Figure 11 is a block diagram that explains the functional structure of the automatic culture device including the culture vessel 1101. is a whole structure diagram in which each constituent element controlled by the control device 1102 possessed by the information processing device is connected to each other or to the incubator 1103. In addition, the above-mentioned closed system culture vessel 1101 is provided in the incubator 1103. The control device 1102 is connected to a temperature adjustment section 1104 for controlling the temperature of the incubator 1103, a gas concentration adjustment section 1106 having a gas supply section 1105 for controlling the gas concentration in the incubator 1103, a pump 1107 provided in a closed system flow path circuit for automatically transferring the culture medium in the closed system culture vessel 1101, and a CO2 / O2 sensor 1108. The control device 1102 is provided in the CPU of a general computer. The control device 1102 causes various programs stored in the storage section to work on the CPU as a processing section. Thereby, the culture environment in the incubator 1103 is controlled by the temperature adjustment section 1104, the gas supply section 1105, the pump 1107, the CO2 / O2 sensor 1108, the gas concentration adjustment section 1106, a temperature sensor 1110, a culture medium container 306, and a culture supernatant container 1111, and it is possible to implement predetermined culture processes in the closed system culture vessel 1101. The exchange of gas in the closed system culture vessel 1101 is performed between the incubator 1103 and the closed system culture vessel 1101 through a gas permeable membrane provided in the closed system culture vessel 1101. In addition, the gas concentration adjustment section 1106 can be directly connected to the closed system culture vessel 1101. The temperature adjustment section 1104, the gas concentration adjustment section 1106, and the CO2 / O2 sensor 1108 can also be connected to the closed system culture vessel 1101. In this structure, gas is directly supplied into the vessel of the closed system culture vessel 1101.

[0058] Figure 12 represents a series of steps for manufacturing a regenerated tissue using the automated culture device having the above functions.

[0059] <Step S1: Flow path setting>

[0060] The automated culture device is started. As information of a plurality of culture devices to be used, a device number or the like is recorded in a job order procedure. An electronic tag or a 2-dimensional bar code is previously provided in the culture device, and the electronic tag or the 2-dimensional bar code is preferably read by a bar code reader or the like, so that it is confirmed in advance that the information of the device number or the like recorded in the job order procedure is consistent. The operator presses and starts an activation switch of an operation section on a control device. In addition, the inside of the device is previously made into a clean environment by sterilization or disinfection.

[0061] Next, it is confirmed in the operation screen of the display of the control section that the internal environment of the automated culture device is appropriate. For example, it is confirmed that the temperature of the incubator is 37°C. These values are not limited, and for example, the temperature can be selected from the range of 0°C to 45°C. In addition, an automated culture schedule is determined in advance by a culture program, and information of the schedule is reflected to the automated culture device. The automated culture schedule also includes a date and time, a liquid amount, and the like of conditions for performing cell inoculation, medium exchange, culture supernatant recovery, inspection tissue recovery, and graft tissue recovery.

[0062] A closed system flow path including a closed system culture container is previously set in the culture device. In addition, at the stage of moving out from a warehouse, an electronic tag or a 2-dimensional bar code previously provided in the closed system flow path is read by a bar code reader or the like, so that it is confirmed in advance that the information of the device number or the like recorded in the job order procedure is consistent. The closed system flow path is constituted by a closed system culture container, a medium container filled with fresh medium, a closed system culture container with a gas permeable membrane for recovering culture supernatant after culture, and the like, and a flow path tube connecting them. The structure corresponds to the kind of the selected closed system flow path. In addition, in the case where the inoculation of cells is performed by manual work, the inoculation is previously performed in a safety cabinet, and a closed system culture container in a state of being filled with a cell suspension is set in the device. In the case where the inoculation of cells is automatically performed by the automated culture device, a cell bottle is previously put into a cell suspension prepared to a predetermined concentration in the safety cabinet, and a closed system culture container including the cell suspension is set in the device. After the start of the automated culture, the automated culture device performs the inoculation by transferring the cell suspension from the cell bottle to each closed system culture container.

[0063] <Step S2: Culture of cells>

[0064] The closed system culture vessel is left at rest for a predetermined time. The culture is maintained at 37°C by the incubator. The air in the device is always stirred by a fan to make the temperature distribution uniform. In addition, in order to improve the manufacturing safety, a particle counter, a viable cell count measuring device can also be installed on the device to monitor the cleanliness. With respect to the gas exchange, in the case of using a closed system culture vessel having a gas permeable membrane, a filter, or the like, the gas phase in the incubator and the gas phase in the closed system culture vessel are exchanged via the gas permeable membrane, the filter, or the like. In the case of using a closed system culture vessel having a gas feeding flow path, the gas exchange is performed by directly feeding a predetermined gas into the closed system culture vessel. The gas exchange is performed several times or more per day during the culture period. As an example of the gas to be supplied, air containing CO2 at a concentration of 5% is used. The gas is sent out from a gas cylinder while controlling the flow rate by a gas flow meter, and is supplied to each closed system culture vessel through a humidifier bottle in order to saturate the water vapor. The unnecessary gas after being fed into the closed system culture vessel is discharged to the outside of the flow path via a filter.

[0065] The filter is preferably a filter of 0.22 μm or more.

[0066] <Step S3: Medium exchange>

[0067] The medium exchange is performed once every several days during the culture period. It can be a medium addition in which only fresh medium is added. The medium stored at 4°C in a refrigeration chamber is sent to a preheating bottle and preheated to 37°C. It can also be warmed to 37°C while being sent at a lower speed. In the medium exchange, first, the old medium is discharged from the closed system culture vessel. After the discharge, the new medium is quickly supplied into the closed system culture vessel. The old medium is finally discharged to a culture supernatant container. The culture supernatant in the culture supernatant container is recovered as needed, and the growth state of the cells is evaluated by a medium component analysis. The medium can also be exchanged by extruding the new medium in a state in which the old medium is added.

[0068] <Step S4: Confirmation of whether it is the day before the transplantation scheduled day>

[0069] It is confirmed whether it is the day before the transplantation scheduled day, and if it is not the day before, the process returns to Step S2 to continue the culture. If it is the day before, the process proceeds to the next step.

[0070] <Step S5: Recovery of the test tissue>

[0071] In the case of culturing a plurality of closed system culture containers at the same time, a part of the closed system culture container can also be recovered for inspection. The door of the culture device is opened, and the flow path tube of the closed system culture container for inspection is aseptically cut off by means of heat sealing or the like, and the closed system culture container is taken out. The closed system culture container that has been removed is carried outside the safety cabinet or CPF, and the inspection is promptly performed. For example, the cell number, survival rate, expression of specific proteins, and the like of the biological test sample are evaluated.

[0072] <Step S6: Culture and medium exchange immediately before transplantation>

[0073] Culture is performed by the same operation as in Step S3. Then, immediately before Step S7 is performed, medium exchange based on the same operation as in Step S4 is performed.

[0074] <Step S7: Recovery and carrying of tissue for transplantation>

[0075] In the case where the evaluation result in Step S5 is determined to be a state suitable for transplantation, the biological test sample is recovered and used for regenerative medical treatment. As in Step S5, the closed system culture container is aseptically separated from the closed system flow path and taken out from the incubator. Carrying treatment into the safety cabinet is performed as necessary.

[0076] The closed system culture container is housed in a transport container for short distance or long distance in the delivery room. By using a heat storage material, an airtight container, packaging, or the like, the effects of temperature, pressure, impact, and the like during transportation are minimized throughout the entire course of transportation. The transport container is transported outside the CPF in this state, and is transported to the operating room by means of a vehicle, a railway, an airplane, a hand-held device, or the like as necessary. Before treatment in the operating room, cell observation based on a microscope is performed as necessary as a reception inspection. In the case of short distance transportation, it is assumed that the state hardly changes compared to immediately before transportation, and therefore it is also possible to perform it by the judgment of the operator.

[0077] <Step S8: Transplantation>

[0078] After arriving at the operating room, the regenerative tissue is taken out of the closed system culture container. At the time of opening, the outside of the closed system culture container can be attached with organisms such as bacteria and particles, and therefore it is aseptically opened in a manner that maintains the cleanliness inside the closed system culture container.

[0079] Finally, the closed system flow path for culture is removed. Next, by appropriately operating the inside of the device, sterilization by purging with a purging gas or wiping with ethanol or the like is performed, and a clean state is formed. The various software of the automatic culture device is ended, and the work of the automatic culture device is ended.

[0080] According to the preferred embodiment of the automatic culture device including the closed system culture container configured as above, for a plurality of culture devices, a high versatility closed system flow path is used, and a culture program using the plurality of culture devices is used, thereby improving the concentration of the culture device, and as a result, the manufacturing cost can be reduced.

[0081] <Embodiment II>

[0082] Using Figure 13 An embodiment using an automatic culture device having a different structure from that of Embodiment 1 will be described.

[0083] In this embodiment, the refrigeration section having in Embodiment I is not provided. Further, the culture medium container and the culture supernatant container are provided on the side surface of the device at room temperature. In Figure 13 The culture medium container 1301 is provided in the

[0084] The closed system flow path can be provided as in Embodiment I. The culture device can be concentrated by a method such as stacking. Further, for a plurality of culture devices, one closed system flow path can be used to culture cells. Thus, a large number of cells can be cultured. In addition, since a plurality of culture devices are used, the culture program shown in Embodiment 1 can be used.

[0085] <Embodiment III>

[0086] Using Figure 14 An embodiment using an automatic culture device having a different structure from that of Embodiment I will be described.

[0087] In this embodiment, the refrigeration section having in Embodiment I is not provided. Further, the flow path mechanism section 102 in Embodiment I is integrated with the incubator 101. The culture medium container, the culture supernatant container, and the like are provided on the side surface of the device at room temperature. In Figure 14 The culture medium container 1401 is provided in the

[0088] The closed system flow path can be provided as in Embodiment I. The culture device can be concentrated by a method such as stacking. For a plurality of culture devices, one closed system flow path can be used to culture cells. Thus, a large number of cells can be cultured. In addition, since a plurality of culture devices are used, the culture program shown in Embodiment I can be used.

[0089] Explanation of Symbols

[0090] 101...incubator

[0091] 102...flow path mechanism section

[0092] 103...refrigeration section

[0093] 104...pump

[0094] 105 solenoid valve

[0095] 106 notch portion

[0096] 107 setting table

[0097] 108 humidification tray

[0098] 201 closed system culture vessel

[0099] 202 culture medium container

[0100] 203 flow path tube

[0101] 204 filter

[0102] 301 filter-external closed system culture vessel

[0103] 302 filter

[0104] 303 flow path tube

[0105] 304 pump

[0106] 305 solenoid valve

[0107] 306 culture medium container

[0108] 307 culture supernatant container

[0109] 308 closed system culture vessel with gas-permeable membrane

[0110] 309 gas-permeable membrane

[0111] 310 closed system culture vessel for perfusion culture

[0112] 1101 closed system culture vessel

[0113] 1102 control device

[0114] 1103 incubator

[0115] 1104 temperature adjustment portion

[0116] 1105 gas supply portion

[0117] 1106 gas concentration adjustment portion

[0118] 1107 pump

[0119] 1108 CO2 / O2 sensor

[0120] 1109... display device

[0121] 1110... temperature sensor

[0122] 1111... culture medium container / culture supernatant container

[0123] 1301... culture medium container

[0124] 1401... culture medium container

Claims

1. A cell culture system provided with an automatic culture device and an information processing device, characterized in that the automatic culture device is provided with: a plurality of closed system flow paths that can be set and detached; and a plurality of culture devices, the plurality of closed system flow paths being configured to enable different culture methods to be performed using the culture devices connected to each closed system flow path, the information processing device is provided with: an input device that accepts, as input, at least one piece of data selected from the group consisting of data of an identifier of a patient, data relating to a transplantation method, data relating to a cell type, data relating to a necessary cell number, and data relating to a treatment plan, in a selection relating to a treatment method for the patient; a calculation device that determines the treatment method based on the input data, generates information relating to cells to be manufactured, and limits and selects, from among options relating to a cell culture method, types and numbers of the culture devices, and types and numbers of the closed system flow paths, the cell culture method, the types and numbers of the culture devices, and the types and numbers of the closed system flow paths to be used, in accordance with the cells to be manufactured; and an output device that outputs the numbers of the closed system flow paths to be used and the numbers of the culture devices to be used, in the information processing device, a program for using the plurality of culture devices is used, the program having an electronic tag that associates each culture device and each closed system flow path with a cell culture method.

2. The cell culture system according to claim 1, characterized in that the closed system flow path is provided with: a valve for opening and closing a flow path tube; a pump for performing liquid or gas transfer of a fluid or gas in the flow path tube; a culture medium container for holding fresh culture medium; and a culture supernatant container for holding culture supernatant after culture.

3. The cell culture system according to claim 1 or 2, characterized in that one closed system flow path is provided for each of the plurality of automatic culture devices.

4. The cell culture system according to claim 1 or 2, characterized in that the output device outputs a job procedure book relating to the closed system flow paths to be used.

5. The cell culture system according to claim 1 or 2, characterized in that the output device outputs a job procedure book relating to the culture devices selected. ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​

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