JP5813680B2 - Cell culture equipment - Google Patents

Description

  The present invention relates to a cell culture apparatus for culturing cells, and more particularly to an automatic culture technique for culturing a large amount of cells efficiently with aseptic cleanness.

  Currently, cell culture work for regenerative medicine is performed by a skilled worker under a strict manufacturing process in a clean room sterilized without limit. In particular, in regenerative medicine such as heart failure treatment using cells collected from muscles, a large amount of cells must be cultured on a culture surface having an area of about 1 tatami per patient. For this reason, there is an increase in the burden on workers and the time and cost required for education and training of workers, human error and sample misconception, and biological contamination (contamination) by humans with bacteria etc. It becomes easy to occur, and a lot of cost is required for these measures. This is a big barrier in the spread and industrialization of cell mass culture. Therefore, it is expected that these problems can be solved by automating a series of culture operations using an instrument.

  However, in the culture treatment of a large amount of cells, since a large amount of the cell suspension is handled in a suspended state that damages the cells, it is necessary to shorten the treatment time as much as possible. Furthermore, a subculture work for re-seeding dense cells into another culture container in a loose state and a concentration work for collecting cells floating in a large amount of medium at the end of the culture are required. In a culture apparatus that cultures a large amount of cells, it is a problem how to satisfy them while maintaining cleanliness.

  In order to solve this problem, for example, Patent Document 1 presents a configuration that can use a pump to move the cell suspension between the processing elements and send the solution. This shortens the distance between the processing elements. Patent Document 2 provides a method of moving a cell suspension to a plurality of processing elements with a single pump. This enables a combination of processing elements and enables efficient cell suspension delivery.

International Publication WO2005-059091 US Publication No. 2011-0207222

  In the automatic culture apparatus as described in Patent Document 1, an independent pump is used for feeding the cell suspension between the processing elements, and the pump installation mechanism and the control mechanism are increased in size. , Accompanied by complication of flow path installation. Accordingly, there is a problem that the larger the number of cells to be processed, the larger and complicated the entire system becomes, and the cost increases.

  In an automatic culture apparatus such as Patent Document 2, the cell suspension and various solutions are sent to each processing element via a pump, but the arrangement of each processing element including the pump is specified. Not. For example, when processing elements with different operating temperature zones such as a medium stored at 4 ° C. and a culture space cultured at 37 ° C. are processed with one pump, the tube between the processing elements becomes long depending on the position of the pump. As a result, the problem is that damage to cells due to temperature and cell suspension state increases. The problem is whether the flow path can be easily installed in the apparatus and the influence on the cells is minimized by defining the optimal installation location of the pump.

  Currently, in regenerative medicine that uses human cells and transplants cells and tissues prepared in culture to humans, the tissues collected in the operating room are placed in aseptically treated test tubes, etc. Necessary cells are isolated from the tissue in a clean room (CPC: Cell Processing Center) compliant with manufacturing standards (GMP: Good Manufacturing Practice), and subjected to the desired adjustment and cultured. In order for the collected cells to be cultured without any contamination within the production process, it must be produced manually in a process and environment that meets particularly strict regulations. Even in an automatic culture apparatus that automatically cultivates cells by a machine, the cells or tissues produced by the apparatus must not be biologically contaminated by any bacteria or viruses in the production process. Under such circumstances, a system has been developed in which a culture vessel itself for automatic culture has a closed structure, the inside of which can be sterilized, and a cell is cultured by supplying driving force from the outside.

  However, in order to culture in a culture vessel having a closed system structure, how to improve the efficiency of the mechanism and control method for performing the culture process is a problem. In particular, in order to process a large amount of cells, further increase in efficiency is required when the culture vessel becomes large and the culture process becomes complicated. In an automatic culture device that has subculture work to replace culture vessels and cell concentration adjustment work (cell concentration), and collects a large amount of cells by expansion culture, in a flow path having each element including a large area culture vessel A configuration that minimizes manufacturing errors and efficiently and safely installs the flow path inside the apparatus is required.

  In view of the above situation, an object of the present invention is to provide a cell culture device capable of performing culture treatment while reducing cell damage.

In the present invention, in order to solve the above problems, a cell culture device,
A pump unit for supplying and discharging cells and medium, a culture container unit for culturing cells by holding cells and medium, a medium storage unit for retaining cells and medium, a cell concentration unit for concentrating cells, a pump unit, , A culture vessel section, a culture medium storage section, and a flow path composed of modules for connecting each of the cell concentration sections and a control section for controlling the pump section, and the culture container section around the pump section A cell culture device having a configuration in which a medium storage unit and a cell concentration unit are arranged is provided.

  According to the configuration of the present invention, it is possible to provide a cell culture device that can shorten the flow path of the entire device and can efficiently culture cells with minimal cell damage.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a front view showing an outline of an example of an automatic culture apparatus according to Example 1 provided with a flow path. It is a front view which shows the outline of the automatic culture apparatus which closed the door based on Example 1. FIG. It is a front view which shows the inside of the automatic culture apparatus which removed the flow path based on Example 1. FIG. It is a block diagram which shows the structure of the flow path which consists of several modules based on Example 1. FIG. It is a block diagram which shows the state before connecting a seal | sticker to the nail | claw of the seal | sticker mechanism based on Example 1. FIG. It is a block diagram which shows the state after connecting a seal | sticker to the nail | claw of a seal mechanism based on Example 1. FIG. 1 is a front view showing a configuration of a transport jig for an automatic culture apparatus according to Example 1. FIG. It is a front view which shows one structure of the conveyance jig which installed the flow path based on Example 1. FIG. 3 is a configuration diagram illustrating an example of an arrangement of culture element devices according to Example 1. FIG. FIG. 6 is a configuration diagram illustrating another example of the arrangement of culture element devices according to the first embodiment. FIG. 6 is a configuration diagram illustrating another example of the arrangement of culture element devices according to the first embodiment. FIG. 6 is a configuration diagram illustrating another example of the arrangement of culture element devices according to the first embodiment. It is a block diagram which shows an example of the control structure of the whole dynamic culture apparatus based on the self-Example 1. FIG.

  Embodiments of the present invention will be described below with reference to FIGS. However, it should be noted that the embodiments described below are merely examples for realizing the present invention and do not limit the technical scope of the present invention. In each drawing, the same reference numerals are assigned to common components.

  In the cell culture device of the present invention, the culture element is arranged around the pump part including the pump and the electromagnetic valve inside the device. A schematic configuration of a preferred embodiment of the cell culture device of the present invention will be described below.

  In order to realize the cell seeding, culture, passage, and cell concentration steps necessary for large-scale cell culture, the cell culture device includes a pump unit, a primary culture container unit, a secondary culture container unit, a medium storage unit, and a cell concentration unit. The five culture elements are configured, and the flow path is configured from five modules correspondingly. First, a pump that feeds each solution such as cell suspension and culture medium, a solenoid valve that defines the feeding destination, and also holds the solution when moving between the culture elements. A pump unit including a tank and the like and a first module were used. And the primary culture container part, the second module, the secondary culture container part, and the third module for the passage to expand the cell culture area are arranged around the pump part and the first module. did.

  The primary culture vessel portion and the secondary culture vessel portion are within 37 ° C., and the inside of the culture vessel maintains a state close to 37 ° C., 5% carbon dioxide concentration, and humidity 100%. After the cells cultured in the primary culture container part are held in the tank in the pump part, the culture surface is sent to a secondary culture container part with a larger area, and subculture is performed, so that more cells can be cultured. Become. Preferably, there are microscopes around the primary culture container part and the secondary culture container part, and the cells inside each culture container can be imaged. Furthermore, a pump storage unit, a first module, and a peripheral module, a medium storage unit for storing each solution such as a cell suspension and a medium, and a waste solution at about 4 ° C., and a fourth module are arranged around the first module. The temperature of the culture medium storage unit is different from that of the primary culture container unit and the secondary culture container unit.

  And when a primary culture container part and a secondary culture container part are arranged in the cell culture apparatus main body, and a medium storage part is arranged outside, a seal mechanism for passing a liquid-feeding tube through the door of the cell culture apparatus main body, etc. Can be divided into areas. Furthermore, when a large amount of cells are used for the next step or treatment after the expansion culture in the secondary culture container portion, it is often necessary to concentrate the cells in the cell suspension by centrifugation or the like. Therefore, a cell concentrating unit and a fifth module for increasing the cell density with the cell suspension are arranged around the pump unit and the first module. The cells cultured in the secondary culture container section are held in a tank in the pump section, and then sent to the cell concentration section to concentrate the cells.

  When a large amount of cell culture is performed, a series of flows in which the cells move through the flow path in the cell culture apparatus are preferably the fourth module, the first module, the second module, the first module, the first module, The third module, the first module, and the fifth module are all in this order, and all the culture elements or movements between the modules are all via the pump unit, the first module, and preferably the tank that is the accumulator inside. did. Therefore, the primary culture container part and the second module, the secondary culture container part and the third module, the medium storage part and the fourth module, the cell concentration part and the fifth module, the pump part and the first module Arranged as center.

  This minimizes the tube length of the channel of each culture element module, shortens the time for feeding liquid that damages cells, minimizes the amount of medium remaining in the tube, and improves the ease of channel installation. It becomes. One pump can perform all processing, simplifying the configuration of related equipment, saving space, and simplifying the control method. Furthermore, the arrangement of the culture elements around the pump part increases the degree of freedom of arrangement, enables the arrangement according to the contents of cell treatment, and has versatility. In addition, the modularization of the flow path makes it possible to manufacture and install the flow path in module units, simplify the flow path configuration, and prevent erroneous connection and space saving.

  Further features of the present invention will become apparent from the following description of various embodiments and the accompanying drawings.

  As Example 1, an example of an automatic culture apparatus capable of improving the efficiency of culturing a large amount of cells will be described.

  First, FIG. 1 which shows one structural example of the automatic culture apparatus according to Example 1, FIG. 2 which shows the overall outline of the state where the door of the automatic culture apparatus is closed, and the whole state where the flow path inside the automatic culture apparatus is removed The overall configuration of the automatic culture apparatus of Example 1 will be described with reference to FIG. 3 showing the outline, FIG. 4 showing the outline of the configuration of the flow path installed in the automatic culture apparatus, and the like.

<Overall configuration of automatic culture device>
The automatic culture apparatus 10 includes a cell culture apparatus body including a cell culture chamber, a medium storage unit, a control unit, and a cell concentration unit as basic components. Each basic configuration will be described below.

  As shown in FIG. 1, the entire automatic culture apparatus 10 includes a cell culture chamber 13 and an automatic culture apparatus main body 11 including a culture element therein, a culture medium storage unit 14, a control unit 15, and a cell concentration unit 16. The cell culture process is performed with the path 40 installed. In the cell culture chamber 13, which is inside the automatic culture apparatus main body 11, various culture elements, that is, a primary culture container part 20 that holds a primary culture container 41 and a secondary culture container part 21 that holds a secondary culture container 42. In addition, a flow path 40 constituted by the tube 45 and a pump unit 31 that enables liquid feeding in the flow path 40 are arranged. The pump unit 31 includes a pump 23, a solenoid valve 24 and a solenoid valve base 25 that enable switching in the flow path 40, a tank 26 that is an accumulator that can hold a cell suspension therein, and a tank in which the tank is installed. A holding unit 27 is arranged. A gas supply unit 22 for supplying a mixed gas into the primary culture vessel 41 and the secondary culture vessel 42 is disposed at the upper part of the automatic culture apparatus main body 11 and connected to the gas connector 28.

  Among the culture elements in the automatic culture apparatus main body 11, the solenoid valve base 25 in the pump unit 31 is detachable from the automatic culture apparatus main body 11 as will be described later. The gas supply unit 22 is for supplying a mixed gas having a 5% carbon dioxide concentration to the atmosphere essential in the cell culture environment. The gas mixed in the gas supply unit 22 is sent to the primary culture container 41 and the secondary culture container 42 in the automatic culture apparatus main body 11 through a gas connector 28 including a filter and a sterile connection connector. The gas supply unit 22 also has a heat exchange function and a humidification function of 37 ° C., for example.

  In the configuration of the present embodiment, the culture medium storage unit 14 installed outside the automatic culture apparatus main body 11 has a refrigerator 29 inside, such as a cell suspension, a culture medium, a washing solution, a cell detachment solution, and a waste solution. The medium element 43 can be stored. The internal temperature of the culture medium storage unit 14 can be maintained at about 4 ° C., for example.

  Similarly, in the cell concentration unit 16 that is installed outside the automatic culture apparatus main body 11 and concentrates the cultured cells, a cell concentration element 44 that can adjust the cell concentration using, for example, a centrifuge or a filtration membrane. Can be held.

The automatic culture device main body 11, the cell concentration unit 16, and the culture medium storage unit 14 each have an automatic culture device main body door 12, a cell concentration unit door 17, and a culture medium storage unit door 18, and can be opened and closed.
FIG. 2 is a state diagram in which all the doors of the automatic culture apparatus 10 of FIG. 1 are closed, and a handle 19 is installed on each door. The automatic culture apparatus 10 is in a cell culture state or a state shown in FIG.

  As will be described later, the control unit 15 of the automatic culture apparatus 10 controls the operation of the automatic culture apparatus main body 11 including the pump unit, the culture medium storage unit 14, and the cell concentration unit 16, and culture based on sensor values and the like. Execute various programs such as schedule management. The control unit 15 includes an input unit 81 and a display unit 80, and enables integrated control of the entire automatic culture apparatus 10.

  Claws are connected between the cell culture chamber 13 and the cell concentrating unit 16 of the automatic culture apparatus main body 11 and between the cell concentrating unit 16 and the medium storage unit 14 via the tube 45 of the flow path 40 so that the insides can be connected. A sealing mechanism including 32 and a seal 33 is configured. Thereby, it becomes possible to hold | maintain the internal temperature of the cell culture chamber 13, the cell concentration part 16, and the culture medium storage part 14, respectively.

  FIG. 5A and FIG. 5B are schematic diagrams relating to a seal mechanism including a seal and a claw between the cell culture chamber 13 and the cell concentration unit 16 and between the cell concentration unit 16 and the medium storage unit 14. By inserting the tube 45 of the flow path 40 into the groove 34 of the seal 33 as shown in FIG. 5A and fitting into the claw 32 as shown in FIG. The space between the cell concentration unit 16 and the medium storage unit 14 is separated, and the medium elements 43 such as the medium, the washing liquid, the cell suspension, and the waste liquid can be fed while maintaining a clean state.

  FIG. 3 shows the arrangement of each culture element as viewed from the front of the automatic culture apparatus 10 in a state where the flow path 40 is removed from the automatic culture apparatus 10 and each door is opened. In the cell culture chamber 13 inside the automatic culture apparatus main body 11, an internal temperature is maintained at 37 ° C., and a primary culture container part 20 and a secondary culture container part 21, which constitute a culture container part, a pump 23, a solenoid valve base An installation unit 30 (including a solenoid valve base 25) and a pump unit 31 including a tank holding unit 27 are arranged. In the primary culture container unit 20, the primary culture container 41 is held, and the primary culture container 41 can be swung and maintained horizontally from the rocking mechanism 46. Moreover, the observation mechanism which abbreviate | omitted illustration is provided, and the cell state inside the primary culture container 41 can be observed.

  Since the secondary culture container 42 of the secondary culture container unit 21 performs expansion culture by passage from the primary culture container 41, the culture area of the secondary culture container 42 is equal to or larger than that of the primary culture container 41. . Therefore, the secondary culture container unit 21 can have a multistage function of the secondary culture container 42 by, for example, a link mechanism or the like. Further, in the secondary culture container section 21, the secondary culture container 42 is held, and the secondary culture container 42 can be swung and maintained horizontally from the rocking mechanism 47. In addition, an observation mechanism (not shown) is provided, and the cell state inside the secondary culture vessel 42 can be observed.

  In the pump unit 31, for example, a mechanism for supplying driving force aseptically from the outside of the tube 45 such as the peristaltic pump 23 is used. Moreover, the rotation of the pump 23 can be switched between forward rotation and reverse rotation, the rotation speed can be changed, and a plurality of tubes 45 can be processed according to each situation. One or a plurality of electromagnetic valves 24 are installed in the electromagnetic valve base 25 and the flow path 40 is switched. As will be described later, the electromagnetic valve base 25 can be detached from the cell culture chamber 13, and the flow path 40 can be installed in the electromagnetic valve 24 outside the automatic culture apparatus 10. As is apparent from the drawing, the electromagnetic valve base installation portion 30 is inside the cell culture chamber 13. The tank holding unit 27 holds the tank 26 that can contain the cell suspension discharged from the primary culture vessel 41 and the secondary culture vessel 42, and has a sensor (not shown) to measure the weight of the tank 26. Is possible.

  The inside of the culture medium storage unit 14 in the present embodiment has a refrigerator 29 structure, and can hold, for example, about 4 ° C. with respect to the culture medium element 43. In addition, a sensor for measuring the weight of the culture medium element 43 can be provided therein to obtain the amount used. The cell concentration unit 16 holds the cell concentration element 44. The temperature inside the cell concentration unit 16 can be maintained from about 4 ° C. to room temperature or 37 ° C. depending on the cells. This prevents cells from adhering inside the cell concentration element 44 and enables temperature buffering when the medium element 44 is fed into the cell culture chamber 13. As the cell concentration element 44, various cell concentration devices that are commercially available can be used.

  FIG. 4 shows the overall arrangement of the flow path 40 of the automatic culture apparatus of this example. Depending on the culture element equipment, the flow path 40 is composed of a plurality of modules. The flow path 40 has a closed system structure, and is mainly composed of a tube 45, a connector, a filter, a check valve, and fittings. Since the structure is complicated, treatment by EOG sterilization (Ethylene Oxide Gas Sterilization), gamma ray sterilization, and electron beam sterilization is desirable. The flow path 40 can be assembled for each module, and the modules can be connected by a sterile connection connector or the like, so that the ease of installation of the flow path 40 can be improved.

  As shown in FIG. 4, the flow path of the automatic culture apparatus 10 of the present embodiment can be divided into five modules surrounded by five broken lines. A flow path component including the primary culture container 41 installed in the primary culture container element unit 20 is referred to as a primary culture container module 35. A flow path component including the secondary culture container 42 installed in the secondary culture container element 21 is referred to as a secondary culture container module 36. The secondary culture container module 36 is equivalent or large in size as compared with the primary culture container module 35 due to expansion culture by passage. A flow path component installed in the tank holding unit 27 is a pump module 37. The pump module 37 is an accumulator that temporarily includes a tube 45 that is adapted to the pump 23 for connection to the pump 23, a tube 45 that is installed in the electromagnetic valve 24, and a cell suspension for installation in the pump unit 31. It consists of a tube connected to the tank 26. The tube 45 for the pump 23 may be branched into a plurality. Furthermore, a heat exchange mechanism such as a heater can be installed in the pump module 37 to warm the culture medium. The tank 26 may be a medium bag or the like that has been processed so that cells do not adhere easily.

  In addition, a flow path component portion that is composed of the cell concentration element 44 and is installed in the cell concentration unit 16 is referred to as a cell concentration module 38. It also includes a medium bag that responds to changes in the amount of solvent due to cell concentration, and a flow path configuration for the pump. A flow path component that is composed of medium elements 43 such as a cell suspension, a culture medium, a washing liquid, a cell detachment liquid, and a waste liquid and is installed in the medium storage unit 14 is referred to as a medium module 39.

  As is clear from FIG. 4, the automatic culture apparatus 10 of this embodiment includes a primary culture container module 35, a secondary culture container module 36, a cell concentration module 38, and a culture medium module 39, with the pump module 37 as the center. Therefore, the degree of freedom of module connection is increased, the module can be arranged according to the cell, and versatility is improved. In addition, it is possible to prevent installation and contamination due to incorrect connection and incorrect installation. In addition, the pump module 37, the primary culture container module 35, the secondary culture container module 36, the culture medium module 39, and the cell concentration module 38 described above are respectively referred to as a first module, a second module, a third module, Sometimes called the fourth module or the fifth module.

That is, the module of the present embodiment includes a first module connected to the accumulator, pump, and solenoid valve, a second module connected to the first culture vessel, and a third module connected to the second culture vessel. It is composed of a module, a fourth module connected to the culture medium storage unit, and a fifth module connected to the cell concentration unit.
With reference to FIGS. 6A and 6B, an example of a method for installing the flow path 40 including a plurality of modules in the automatic culture apparatus 10 using the transport jig 50 will be described. As shown in FIG. 6A, the transport jig 50 is configured by a frame for installing the flow path 40. The first frame 51 in which the primary culture container module 35 described above is installed, the second frame 52 in which the secondary culture module 36 is installed, the pump module 37 including the electromagnetic valve base 25, the cell concentration module 38, and the culture module 39 are provided. It has the 3rd flame | frame 53 to install.

  One or more secondary culture vessels 42 can be held in the second frame 52. When a heavy object is placed on the first frame 51, the second frame 52, and the third frame 53, wheels and counterweights are installed on the base 54 in order to make conveyance easy and stable. The first frame 51 and the second frame 52 are attached with a mechanism (not shown) that can move up and down 55. In addition, the third frame 53 can be moved up and down 55 as required.

  FIG. 6B is a diagram illustrating a state where the flow path 40 is installed in the transport jig 50 of the present embodiment. The culture medium module 39 can be suspended on the third frame 53 as required. The state in which the transport jig 50 in the state of FIG. 6B is moved to the automatic culture apparatus 10 in the state shown in FIG. Each door is closed and the cell culture process is performed in the state shown in FIG. The transport jig 50 enables a series of operations to install the flow path 40 in the automatic culture apparatus 10 easily and accurately even with the configuration of the enlarged flow path 40.

<Arrangement configuration of culture elements including modules>
Then, the installation arrangement | positioning of the culture | cultivation element of the automatic culture apparatus 10 of a present Example and the modularized flow path is demonstrated using FIG. 7A-FIG. 7D. In the configuration of the present embodiment, versatility can be realized in the installation of the flow path 40 in the automatic culture apparatus 10 from the modularization of the flow path 40 around the pump 23 and the arrangement of the culture element devices corresponding thereto.

  FIG. 7A shows an example of the arrangement of the culture element devices according to the first embodiment described in the figure, and FIGS. 7B, 7C, and 7D show modified embodiments of the arrangement of the culture element devices. In the arrangement configuration of each figure, the primary culture container section 20, the secondary culture container section 21, the cell concentration section 16, and the culture medium storage section 14 are connected to the pump section 31 with the pump section 31 as the center. As a result, each connection can be minimized. In addition, each module 35-39 of the flow path 40 demonstrated in FIG. 4 shall respond | correspond to arrangement | positioning of each culture | cultivation element apparatus. In particular, the insides of the primary culture vessel portion 20 and the secondary culture vessel portion 21 maintain a state close to 37 ° C., 5% carbon dioxide concentration, and humidity 100%. Moreover, the inside of the culture medium storage unit 14 is set to 4 ° C., for example.

  7A, the cell culture chamber 13 inside the automatic culture apparatus main body 11 has a primary culture container section 20, a secondary culture container section 21, and a pump section 31, and the cell concentration section 16 outside the automatic culture apparatus main body 11 is used. There is a medium storage unit 14. In the figure, the flow path 40 is directly connected from the pump unit 31 to the cell concentration unit 16 and the culture medium storage unit 14, but the flow path 40 to the culture medium storage unit 14 may pass through a part of the cell concentration unit 16. Including.

  In this configuration, for example, the temperature inside the cell culture chamber 13 is 37 ° C., and the outside of the cell concentrating unit 16 and the automatic culture apparatus main body 11 is room temperature (assuming around 20 ° C.). In addition, this configuration realizes the steps of cell seeding, culture, passage, and cell concentration necessary for mass cell culture. In cell seeding in the primary culture container 41 of the primary culture container unit 20, the cell suspension in the medium storage unit 14 is fed to the primary culture container unit 20 via the pump unit 31 and cultured. The cell concentration can be changed using a medium or the like. At this time, since the room temperature outside the cell concentrating portion 16 or the automatic culture apparatus main body 11 passes, the temperature rise can be buffered and the damage to the cells can be minimized. Moreover, the distance of the tube 45 which comprises the path | route 40 can be shortened, liquid feeding time can be shortened, and the cell damage in a floating state can be minimized.

  7A, the cells cultured in the primary culture container 41 of the primary culture container unit 20 are suspended and moved to the tank 26 of the pump unit 31 for the passage work, and then the secondary culture container The solution is fed to the secondary culture container 42 of the unit 21. The cell concentration can be changed (other than cell concentration) by detaching the cell with a cell detachment solution or the like with a medium. At this time, since the cell suspension moves at the same temperature in the cell culture chamber 13, there is no cell damage due to temperature. Further, the tube 45 distance can be shortened to shorten the liquid feeding time, and the cell damage in the floating state can be minimized.

  Next, in the medium exchange at the time of culture, the waste liquid discharged from the primary culture container unit 20 and the secondary culture container unit 21 moves to the medium storage unit 14 via the pump unit 31. Then, the medium or the like is fed from the medium storage unit 14 to the primary culture container unit 20 and the secondary culture container unit 21 via the pump unit 31. The cells cultured in the secondary culture container 42 of the secondary culture container unit 21 are put into a suspension state, moved to the tank 26 in the pump unit 31, and fed to the cell concentration unit 16. Cell detachment can be performed with cell detachment solution, and cell concentration can be changed with medium or the like (change other than cell concentration).

  Cell concentration is performed by the cell concentration element 44 of the cell concentration unit 16. In order to prevent cell adhesion in the cell concentration element 44 and minimize cell damage, cell concentration near room temperature is performed. After the cell concentration is completed, the cell suspension is stored in a medium bag or the like installed in the cell concentration unit 16. The cell suspension may be stored in the culture medium storage section 14 by feeding or transporting the liquid from the cell concentration section 16 to the culture medium storage section 14.

  The example process in FIG. 7A described above is summarized with a focus on the movement of the cell suspension. Medium storage unit 14 → pump unit 31 → primary culture container unit 20 → pump unit 31 → secondary culture container unit 21 → pump Part 31 → cell concentrating part 16, and further → medium storage part 14. Here, → indicates the moving direction of the cell suspension.

  As described above, in the automatic culture apparatus 10 of the present embodiment, since the pump unit 31 is interposed during the movement between the culture elements, the primary culture container unit 20 and the secondary culture unit 20 are arranged around the pump unit 31. By arranging the culture container part 21, the culture medium storage part 14, and the cell concentration part 16, the length of the tube 45 of the flow path 40 between each culture element or module is minimized, and the liquid feeding time for damaging the cells is shortened. In addition, it is possible to minimize the amount of medium remaining in the tube 45 and improve the ease of installation of the flow path 40. One pump 23 can perform all processing, simplifying the configuration of related equipment, saving space, and simplifying the control method. In addition, the arrangement of the culture elements around the pump 23 increases the degree of freedom of arrangement, enables arrangement according to the contents of cell treatment, and versatility.

  The arrangement of the culture elements according to other modified examples will be described with reference to FIGS. 7B to 7D. Also in each modification shown below, the flow path can be shortened as a whole apparatus by arranging the culture vessel parts 20 and 21, the culture medium storage part 14, and the cell concentration part 16 around the pump part 31 as a center. It becomes composition.

  About FIG. 7B, the case where it comprises except the culture medium storage part 14 in the automatic culture apparatus main body 11 is shown. Compared with the arrangement configuration of FIG. 7A, the arrangement of FIG. 7B can further save space. In this arrangement configuration, the cell concentration unit 16 is installed in the cell culture chamber 13 of the automatic culture apparatus main body 11. That is, by installing the pump unit 31, the cell concentrating unit 16, the primary culture container unit 20, and the secondary culture container unit 21 in a 37 ° C. environment in the cell culture chamber 13, the cells can be applied to cells in all steps after seeding. Temperature damage can be eliminated, and this is effective for culturing that prevents the influence of temperature on cells. However, if necessary, the pump unit 31 and the cell concentration unit 16 can be installed in the cell culture chamber 13 with different temperature zones.

  7C shows a case where the primary culture container part 20 of FIG. 7B is incorporated into a part of the secondary culture container part 21. FIG. In this case, in the arrangement configuration of FIG. 7B, it is assumed that space is saved by using a partial area of the secondary culture vessel 42 as the primary culture vessel 41. Furthermore, by using the flow path 40 of the primary culture container part 20 and the pump part 31 together with the flow path 40 of the secondary culture container part 21 and the pump part 31, it is possible to further reduce the space.

  The arrangement configuration of FIG. 7D shows a case where all the culture elements are arranged inside the automatic culture apparatus main body 11, and the most space saving can be achieved. However, in order to enclose regions of different temperature zones in which the culture medium storage unit 14 is installed in the automatic culture apparatus main body 11, for example, as shown in FIG. 7D, the primary culture vessel unit 20 and the secondary culture vessel unit 21 are further required. Accordingly, the cell concentration unit 16 and the pump unit 31 are adjacent to each other, and the culture medium storage unit 14 is provided adjacent to the pump unit 31.

<Control system configuration of automatic culture device>
FIG. 8 is a block diagram showing an example of the configuration of a control system such as the control unit 15 for controlling internal devices in the automatic culture apparatus 10. The configuration of this control system can be used in all the device configurations of the above-described embodiments and various modifications.

  In the figure, the configuration of the control system of the automatic culture apparatus 10 includes an input unit (keyboard, mouse, etc.) 81 for inputting data and instructions, a pump unit 31 in the automatic culture apparatus 10, and swing mechanisms 46, 47. The operation control unit 82 for controlling each operation of the control target device in the device, the environment holding device, etc., the display unit 80 showing the control status to the user, and the operation program, parameters, etc. of the control unit 82 are stored. ROM 85, a RAM 86 for temporarily storing data and processing results, a memory 85 for performing operations such as a cache, a heater, a fan, carbon dioxide supply, water supply, etc. An environment holding device 87 in the culture chamber 13, a cell concentration unit environment control unit 88 that controls the environment of the cell concentration unit 16, and a medium storage unit environment control unit 89 that controls the environment of the medium storage unit 14. It is provided.

  In addition, the operation control unit 82 can be further collectively referred to as a control unit by the cell concentration unit environment control unit 88 and the culture medium storage unit environment control unit 89, and at least one central processing unit (CPU: Central Processing Unit). It is configurable. In addition, the control unit can include the memory 83, the communication unit 84, the ROM 85, and the RAM 86 in FIG. 8 and correspond to the control unit 15 of the automatic culture apparatus 10 shown in FIG. Needless to say, the control unit 15, the display unit 80, and the input unit 81 can be configured by a personal computer or the like.

  When the user instructs the culture process to be processed from the input unit 81 or the communication unit 84 by the automatic culture apparatus 10 of the present embodiment described above, the operation control unit 82 sets the initial value according to the culture preparation program stored in the ROM 85. The control of the swing mechanism 46 of the primary culture container section 20 related to the dispensing, the control of the swing mechanism 47 of the secondary culture container section 21, the control of the pump section 31 that controls the pump 23, the electromagnetic valve 24, etc. The display unit 80 notifies the installation of the flow path 40 and detects the installation. Then, the operation control unit 82 controls the swing mechanism 46 of the primary culture vessel unit 20, the control of the swing mechanism 47 of the secondary culture vessel unit 21, the pump 23 and the electromagnetic valve according to the automatic culture program stored in the ROM 85. Control of the pump unit 31 that controls 24 and the like is performed, and cell culture processing is performed. Moreover, the processing status can be shown to the user at any time by the display unit 80 and the communication unit 84. When the cell culture process ends, the display unit 80 and the communication unit 84 indicate the end to the user, and the operation control unit 82 performs the end process according to the end program stored in the ROM 85. As described above, a series of cell culture processes by the automatic culture apparatus 10 can be realized.

  With the configurations of the various embodiments described above, it is easy to install a channel having a culture vessel, prevents mistakes during channel manufacturing and installation, and is efficient with aseptic cleanliness and minimal cell damage. Enables cell culture equipment to be well cultured.

  Note that the present invention is not limited to the above-described embodiments and modifications, and includes various modifications. For example, the above-described embodiments have been described in detail for better understanding of the present invention, and are not necessarily limited to those having all the configurations described. That is, it is possible to add, delete, and replace other configurations for a part of the configuration of the embodiment.

  In addition, each of the above-described configurations, functions, processing units, and the like has been described as an example of creating a program operation that realizes part or all of them by the operation control unit. Needless to say, it may be realized by hardware.

DESCRIPTION OF SYMBOLS 10 Automatic culture apparatus 11 Automatic beauty apparatus main body 12 Automatic culture apparatus main body door 13 Cell culture room 14 Medium storage part 15 Control part 16 Cell concentration part 17 Cell concentration part door 18 Medium storage part door 19 Handle 20 Primary culture container part 21 Secondary Culture container part 22 Gas supply part 23 Pump 24 Electromagnetic valve 25 Electromagnetic valve base 26 Tank 27 Tank holding part 28 Gas connector 29 Refrigerator 30 Electromagnetic valve base installation part 31 Pump part 32 Claw 33 Seal 34 Groove 35 Primary culture container module 36 Secondary Culture container module 37 Pump module 38 Cell concentration module 39 Medium module 40 Channel 41 Primary culture container 42 Secondary culture container 43 Medium element 44 Cell concentration element 45 Tube 46 Oscillation mechanism (primary culture container)
47 Oscillation mechanism (secondary culture vessel)
50 Transport jig 51 First frame 52 Second frame 53 Third frame 54 Base 55 Vertical movement 80 Display unit 81 Input unit 82 Operation control unit 83 Memory 84 Communication unit 85 ROM
86 RAM
87 Environment holding device 88 Cell concentration unit Environment control unit 89 Medium storage unit Environment control unit

Claims (12)

A cell culture device,
A pump unit for holding a pump for supplying and discharging cells and medium;
A first culture vessel portion and a second culture vessel portion for culturing the cells while holding the cells and the medium;
A medium storage unit for holding the cells and the medium;
A cell concentration unit for concentrating the cells;
A flow path constituted by a plurality of modules connecting the pump unit, the first culture container unit, the second culture container unit, the medium storage unit, and the cell concentration unit;
A control unit for controlling the pump unit,
Of the two sides of the pump, the second culture container part is disposed on one side, and at least one of the medium storage part or the cell concentration part is disposed on the other side,
The first culture vessel part is disposed on the upper side of the pump.
A cell culture device. The cell culture device according to claim 1,
Outside the medium storage unit , a housing in which at least the pump unit, the first culture container unit, and the second culture container unit are arranged is disposed.
A cell culture device. The cell culture device according to claim 2,
The cell concentrating portion is disposed outside the housing;
A cell culture device. The cell culture device according to claim 1,
The second culture vessel section cultures the cells cultured in the first culture vessel section;
A cell culture device. The cell culture device according to claim 4,
The cells and the medium are moved in order of the medium storage unit, the first culture container unit, the second culture container unit, and the cell concentration unit through the pump unit via the channel.
A cell culture device. A cell culture device,
A pump unit for holding a pump for supplying and discharging cells and medium;
A first culture vessel portion and a second culture vessel portion for culturing the cells while holding the cells and the medium;
A medium storage unit for holding the cells and the medium;
A cell concentration unit for concentrating the cells;
A flow path constituted by a plurality of modules connecting the pump unit, the first culture container unit, the second culture container unit, the medium storage unit, and the cell concentration unit;
A control unit for controlling the pump unit,
Of the two sides of the pump, the second culture container part is disposed on one side, the medium storage part is disposed on the other side, and on the upper side of the pump or the upper side of the second culture container part, A first culture vessel portion is disposed;
A cell culture device. The cell culture device according to claim 6,
Outside the medium holding part , a housing in which at least the pump part, the first culture container part and the second culture container part are arranged is disposed.
A cell culture device. The cell culture device according to claim 6 or 7,
The cell concentration unit is disposed on the lower side of the pump,
A cell culture device. The cell culture device according to claim 6,
The second culture vessel section cultures the cells cultured in the first culture vessel section;
A cell culture device. The cell culture device according to claim 9,
The cells and the medium are moved in order of the medium storage unit, the first culture container unit, the second culture container unit, and the cell concentration unit through the pump unit via the channel.
A cell culture device. A cell culture device,
A pump unit for holding a pump for supplying and discharging cells and medium;
A first culture vessel portion and a second culture vessel portion for culturing the cells while holding the cells and the medium;
A medium storage unit for holding the cells and the medium;
A cell concentration unit for concentrating the cells;
A flow path constituted by a plurality of modules connecting the pump unit, the first culture container unit, the second culture container unit, the medium storage unit, and the cell concentration unit;
A control unit for controlling the pump unit,
The second culture vessel section is disposed on the side of the pump, the cell concentration section is disposed on the upper side of the pump, the medium storage section is disposed on the lower side of the pump, and the second culture container section On the upper side, the first culture container part is disposed.
A cell culture device. The cell culture device according to claim 11,
The cells and the medium are moved in order of the medium storage unit, the first culture container unit, the second culture container unit, and the cell concentration unit via the pump unit via the flow path,
The second culture vessel section cultures the cells cultured in the first culture vessel section;
A cell culture device.

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