JP2013031461A - Automated culturing apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To perform a culture treatment by: forming a culture container or a flow path into a sealed structure; supplying a drive force; and antiseptically inserting a culture medium or a cell into a culture container of automatic culture system.SOLUTION: A base 18 is assembled under a sterile environment, and a culture medium or a cell is inserted in a sealed flow path 27. A culture container 20 or the sealed flow path 27 is arranged at a most appropriate position by a guide, the base 18 is easily connected to a drive part 19 inside an automatic culture apparatus 10, and a drive force is supplied to the flow path 27. There is provided an automated culture apparatus including easiness in assembling the sealed flow path 27 by a sealed culture container 20 by a base 18; and efficiently culturing a cell with antiseptic cleanliness at GMP level.

Description

  The present invention relates to an automatic culture apparatus for culturing cells, and more particularly to an automatic culture technique for efficiently culturing cells with aseptic cleanliness.

  Conventionally, cell culture work has been performed manually by skilled workers under a strict manufacturing process in a clean room that is sterilized without limit. Therefore, when cultivating a large amount of cells for industrialization, the burden on the worker is increased and the time and cost required for education and training of the worker, human error and sample misconception, and further possessing bacteria etc. There is a possibility that contamination from human beings may occur, and the countermeasures are costly. This becomes a big wall in the industrialization of cell mass culture. Therefore, it is expected to solve these problems by automating a series of culture operations using an apparatus. In view of this, an automatic culture apparatus that mimics the culture work performed manually by using an arm type robot manipulator is mainly developed at present. In such a culture apparatus, how to handle the culture container aseptically using an apparatus is a problem.

  In order to solve this problem, for example, Patent Document 1 provides an example of processing a culture container transfer operation, a medium exchange, and the like using an articulated robot manipulator. In this articulated robot manipulator, it is possible to sterilize itself.

  Further, as disclosed in Patent Document 2, a method of treating the culture vessel and the flow path itself in a closed system is conceivable. This is a device that cultivates chondrocytes under high pressure. The culture vessel and flow channel are sealed, and the culture vessel and partly plugged flow channel are taken out so as not to touch the outside air after the culture is completed. Means for recovering aseptically generated cartilage tissue.

JP 2006-149268 A Japanese Patent Laid-Open No. 2002-315566

  In the automatic culture apparatus as described in Patent Document 1, since the culture container itself is not a closed system, an operation for opening the inside of the culture container by opening the lid is necessary, and the cleanliness inside the automatic culture apparatus must be maintained. However, although not as much as when working manually, there is a problem that the whole system becomes large and costs high.

  In addition, since the automatic culture apparatus such as Patent Document 2 is a closed system, the flow path is attached while maintaining the sterility of the interior, and the medium (culture solution) and cells (cell suspension) are attached. The problem is whether to inject into the flow path. Moreover, when carrying out complicated culture work, it is a problem whether the sealed flow path can be easily installed in the apparatus and the driving force is efficiently applied.

  Currently, in regenerative medicine using human cells and transplanting cells and tissues prepared in culture to humans, the tissues collected in the operating room are placed in aseptically treated test tubes, etc. The tissue is made into cells in a CPC (Cell Processing Center), which is a clean room compliant with (Good Manufacturing Practice), and cultivated after adjusting the target. In order for the harvested cells to be cultured without any contamination, they must be produced manually in a process and environment that meets particularly strict regulations. Even in an automatic culture apparatus that automatically performs cell culture processing by a machine, the cells or tissues produced by the apparatus must not be contaminated with any bacteria or viruses during the production.

  Under such circumstances, when considering the system of the automatic culture apparatus using the above-described open culture container, in order to sterilize the automatic culture apparatus interior and to maintain a highly clean environment inside, Huge air-conditioning equipment and sterilization equipment are required, which may increase the cost and maintenance cost of the equipment itself. Further, during sterilization, the motor drive system is not able to withstand the treatment as it is. Therefore, it is preferable that the culture vessel itself for automatic culture has a sealed structure, the inside is sterilized, and a system for culturing cells by supplying a driving force from the outside is preferable. However, in order to culture in a closed culture vessel, how to put the culture medium and cells in the culture vessel aseptically and carry out the culture treatment becomes a problem.

  The present invention has been made in view of such a situation, and an object thereof is to provide an automatic culture apparatus and culture container installation method aiming at GMP compliance, which can make a tissue using human cells. To do.

  In order to achieve the above object, in the present invention, there is provided an automatic culture apparatus for culturing cells, comprising a culture vessel, a channel connected to the culture vessel, and a base on which the culture vessel and the channel are placed. And an automatic culture apparatus having a base on which a base is mounted and which gives a driving force to the base.

  In order to achieve the above object, according to the present invention, an automatic culture apparatus for culturing cells, which performs cell culture using a heat-retaining part that keeps cells and a culture solution at a predetermined temperature, and a culture vessel A cell culture unit, an air circulation unit that circulates air between the cell culture unit and the heat insulation unit, and a control unit that controls the heat insulation unit, the cell culture unit, and the air circulation unit. Provided is an automatic culture apparatus having a configuration in which a base on which a flow path connected to a container is placed is provided and a drive unit that applies a driving force to the installed base.

  Furthermore, in order to achieve the above object, the present invention provides a culture container installation method for an automatic culture apparatus that performs cell culture using a culture container, wherein the culture container and a flow path connected to the culture container are mounted. After placing the base with the culture vessel and the flow path connected to the base set that is mechanically detachable, move the base set into the automatic culture device and move the base to a predetermined position in the culture unit. A method for installing a culture vessel to be placed is provided.

  That is, in order to achieve the above object, the automatic culture apparatus of the present invention uses a mechanism called a base. This base makes it possible to optimally arrange the culture vessel and the closed system flow path, and easily connect with the drive unit inside the automatic culture apparatus to supply the drive force to the flow path.

  According to the automatic culture apparatus of the present invention, the degree of cleanliness can be maintained at a GMP level capable of culturing human cells, and efficient automatic culture of cells can be achieved by a closed culture vessel or a closed flow path.

It is a figure which shows the whole schematic structure of the automatic culture apparatus which installed the base of the 1st Example of this invention. It is a figure which shows the external appearance of the automatic culture apparatus which closed the door based on a 1st Example. It is a figure which shows the state inside the automatic culture apparatus which removed the base based on a 1st Example. It is a block diagram which shows the state before the connection of a base and a flow path based on a 1st Example. It is a block diagram which shows the state after the connection of a base and a flow path based on a 1st Example. It is a block diagram which shows the state before installing the seal | sticker of a cell culture room and a refrigerator based on a 1st Example. It is a block diagram which shows the state after installing the seal | sticker of a cell culture room and a refrigerator based on a 1st Example. It is a block diagram which shows the state before a base set and the connection with a base based on a 1st Example. It is a block diagram which shows the state after a base set connects with a base based on 1st Example. It is a block diagram which shows the state which a base set inserts a base into an automatic culture apparatus based on a 1st Example. It is a block diagram which shows the structure of the conveying tool which conveys a base set based on a 1st Example. It is a side view showing the state before putting a base in a base set concerning the 1st example. It is a side view showing the state after putting a base in a base set concerning the 1st example. It is a side view which shows the state which inserted the base into the drive mechanism based on 1st Example. It is a side view which shows the state which removed the nail | claw in order to remove a base from a base set based on a 1st Example. It is a side view which shows the state after removing a base from a base set based on a 1st Example. It is a side view which shows the state before connecting the three-way valve and drive mechanism (motor) based on a 1st Example. It is a side view which shows the state before a 3 way valve and a motor connect based on a 1st Example. It is a side view which shows the state after connecting a base, a 3 way valve, and a motor based on a 1st Example. It is a side view which shows the state which switched the three-way valve of the base by the motor based on a 1st Example. It is a side view which shows the state before the drive mechanism (solenoid) connects with the three-way valve which the tube of the base connected to the 1st Example. It is a side view which shows the state which switched the three-way valve of the base by the solenoid based on a 1st Example. It is a figure which shows the automatic culture apparatus front structure before connecting the base with a flow path based on a 1st Example. It is a figure which shows the front structure of an automatic culture apparatus after connecting the base with the flow path based on a 1st Example. It is a figure which shows the automatic culture apparatus front structure at the time of inject | pouring a liquid with a culture container vertical | vertical based on a 1st Example. It is a figure which shows the front structure of the automatic culture apparatus at the time of discharging | emitting a liquid by making a culture container perpendicular | vertical based on a 1st Example. It is a block diagram which shows the structure of control of the whole automatic culture apparatus based on a 1st Example. It is a figure which shows the whole schematic structure of the automatic culture apparatus which installed the base which included the drive system based on a 2nd Example. It is a figure which shows the whole schematic structure of the automatic culture apparatus which has not installed the base which included the drive system based on a 2nd Example. It is a block diagram which shows the state before the connection of the base and the flow path which included the drive system based on a 2nd Example. It is a block diagram which shows the state after the connection of the base and flow path which included the drive system based on a 2nd Example. It is a side view which shows the state before a drive mechanism (solenoid) operate | moves with the three-way valve to which the tube of the base which included the drive system based on the 2nd Example was connected. It is a side view which shows the state which switched the three-way valve of the base which included the drive system by the operated solenoid based on a 2nd Example.

  Prior to describing various embodiments, a preferred configuration of an automatic culture apparatus using the base of the present invention will be outlined below.

  First, in the culture preparation stage, the automatic culture apparatus without a base can be sterilized with a sterilization gas or the like. Thereafter, the environmental maintenance mechanism inside the automatic culture apparatus operates, the refrigerator functioning as a heat retaining unit maintains approximately 4 ° C., and the inside of the cell culture chamber, which is the culture unit, is 37 ° C., 5% carbon dioxide, and humidity 100%. Hold. At the same time, a closed flow path including a closed culture vessel that has been sterilized, a sterilized base capable of installing the culture vessel and the flow path, and a transport means for installing the base in an automatic culture apparatus The base set is equipped with them in a highly clean environment with very few particles and bacteria. A guide or the like can be installed on the base of the flow path to make it easy to understand visually. Among these, the cell suspension and the culture medium can also be installed in the flow path. Thus, the channel can be kept in a sealed structure with the cell suspension and the medium added. Once everything is set up on the base set, move it to or out of the clean room and open the door. A base set is used to connect the base to the drive mechanism. Remove the base from the base set by the base set mechanism. Here, the base and the drive mechanism are connected to enable driving of a three-way valve or a syringe pump. Alternatively, a base configuration including the drive mechanism without being separated from the drive mechanism can be used.

  In this situation, for example, using a syringe pump or the like of the flow path drive mechanism, the cells (cell suspension) and the culture medium (culture solution) are moved from the refrigerator to the tank in the flow path base, and the temperature is set to 37 ° C. To do. At that time, a seal is provided between the cell culture chamber and the refrigerator having different temperature zones and humidity, and the flow path is installed in the through hole of the seal. A flow path is controlled by a flow path drive mechanism and a flow path base, a culture container drive mechanism and a culture container base with a three-way valve or a syringe pump, and a cell suspension or a medium is supplied into the culture container. At that time, the culture vessel is rotated (clockwise as viewed from the front of the automatic culture apparatus) by the rotary shaft motor and is vertically set, and the cell suspension or the medium is supplied from the lower part of the culture vessel. When changing the culture medium, rotate the culture vessel with the rotary shaft motor (counterclockwise when viewed from the front of the automatic culture device) and stand vertically to discharge the cell suspension or culture medium from the bottom of the culture vessel and inspect it Move to the device. Then, the culture vessel is quickly rotated by a rotating shaft motor (clockwise as viewed from the front of the automatic culture apparatus) and vertically placed, and a cell suspension or a medium is supplied from the lower part of the culture vessel. The liquid flow can be efficiently exchanged while keeping the cleanness in one-way. During the culture, the state of the cells in the culture container can be observed with a separately installed microscope or the like. After the predetermined treatment, when the cells grow or become tissues, the base set is used or the culture vessel is taken out alone and used for a desired use. The channel is discarded according to the application, and the base set and the base are sterilized by an autoclave or the like and reused, and the inside of the automatic culture apparatus is sterilized with a sterilizing gas or the like and is used again.

  Subsequently, various embodiments will be described with reference to the accompanying drawings. However, it should be noted that the embodiment described below is merely an example for realizing the present invention and does not limit the technical scope of the present invention. In each drawing, the same reference numerals are assigned to common components.

  1 to 9 are diagrams for explaining the automatic culture apparatus according to the first embodiment. FIG. 1 is an overall schematic diagram of an automatic culture apparatus 10 according to the first embodiment. FIG. 2 is a schematic view of the appearance of the automatic culture apparatus 10 when the cell culture chamber door 11 and the refrigerator door 12 are closed. FIG. 3 is an overall schematic diagram of the automatic culture apparatus 10 when the flow channel base 51 in the automatic culture apparatus 10 is removed. 4A to 4D are schematic diagrams relating to the attachment and detachment of the flow path 27 and the base 18. 5A to 5D are schematic diagrams relating to the base set 50. 6A to 6E are side views regarding the processing of the base 18 by the base set 50. 7A to 7D are side views regarding the connection and operation of the valve 24 and the drive unit 19 of the base 18. 8A to 8D are front views relating to the liquid injection operation to the culture vessel 20 by the rotation mechanism 17. FIG. 9 is a block diagram showing a circuit for operating the automatic culture apparatus 10.

<Configuration of automatic culture device>
The whole structure of the automatic culture apparatus 10 is demonstrated using FIG.1, FIG2 and FIG.3. The automatic culture apparatus 10 includes, as basic components, a cell culture chamber 13 that is a culture unit, a refrigerator 14 that functions as a heat retaining unit, a control unit 15, and a clean air circulation unit 16 that is an air circulation unit. Each basic configuration will be described below.

  As shown in FIG. 1, the entire automatic culture apparatus 10 is composed of a cell culture chamber 13, a refrigerator 14, a control unit 15, and a clean air circulation unit 16, and the cell culture chamber door 11 and the refrigerator door 12 are arranged. By opening it, it is possible to access the inside of the automatic culture apparatus 10. In the cell culture chamber 13, a flow path 27 is provided by a culture container base 21 having a culture container 20, a culture container drive unit 22 connected to the culture container 20, a rotating mechanism 17 that rotates them, a motor 23, a valve 24, a motor 25, and a tank 26. A flow path base 28 to be configured, a flow path drive unit 29 connected thereto, a carbon dioxide supply mechanism 30 for adjusting the environment in the cell culture chamber 13, a carbon dioxide sensor 31, a humidity adjustment mechanism with a temperature sensor 32, a humidity sensor 33, and a fan 34.

  As shown in FIG. 1, the culture vessel base 21, the flow path base 28, and the culture medium base 35 are collectively referred to as a base 18, and the culture container drive unit 22 and the flow path drive unit 29 are collectively referred to as the drive unit 19. Called.

  At the time of cell culture, the inside of the cell culture chamber 13 maintains an environment of, for example, a temperature of 37 ° C., a humidity of 100%, and 5% carbon dioxide. The medium base 21 can be placed in the refrigerator 14, and the internal temperature can be maintained by the seal 46 while the flow path 27 is taken out of the refrigerator 12 by the claw portion 36 attached to the refrigerator. The clean air circulation unit 16 warms the air in the cell culture chamber 13 that has exited from the exhaust port 37 with a heater 39 and supplies it to the cell culture chamber 13 through a clean filter such as a HEPA (High Efficiency Particulate Air) filter with a fan 38. The cleanliness inside the cell culture chamber 13 is maintained. The control unit 15 is independent from other compartments, and blocks the temperature, humidity, and carbon dioxide in the cell culture chamber 13.

  FIG. 2 shows a state of the automatic culture apparatus 10 with the cell culture chamber door 11 and the refrigerator door 12 closed. The automatic culture apparatus 10 is configured to maintain an internal sealing property and to maintain an internal temperature by a heat insulating mechanism. Furthermore, the cell culture chamber door 11 and the refrigerator door 12 are each provided with a stopper and a heat insulation mechanism that are closed when they are closed, and when the doors are closed, the internal sealing and temperature of the automatic culture apparatus 10 are maintained. Can do. The control unit 15 includes a display unit 42 for displaying a status, an input unit 43 for performing operations related to control, and a communication unit 44 that can access an external device such as a PC or a recording device and an internal control device. .

  Further, the claw portions 36-1 and 36-2 of the refrigerator door 12 communicate with the cell culture chamber 13 in the automatic culture apparatus 10 with the refrigerator door 12 closed when the flow path 27 is not attached. Therefore, it is possible to sterilize the cell culture chamber 13, the refrigerator 14, and the clean air circulation unit 16 at a time by supplying a sterilization gas from the sterilization gas supply port 40 with the cell culture chamber door 11 and the refrigerator door 12 closed. It is. Here, since the apparatus is sealed, the sterilization gas does not leak to the outside of the automatic culture apparatus 10. At this time, the sterilization gas does not enter the control unit 15. After the completion, after the sterilizing gas remaining from the sterilizing gas exhaust port 41 is discharged, aeration is performed for a predetermined time to prepare for the next culture.

  As shown in FIG. 3, the flow path 27 in the cell culture chamber 13 and the refrigerator 14, the culture container base 21, the flow path base 28, and the culture medium base 35 are respectively a culture container drive unit 22 and a flow path drive unit. 29. It can be removed from the refrigerator 14, and after the cell culture is completed, the inside is put in this state, and the sterilization process described with reference to FIG. 2 is performed. The culture vessel drive unit 22 and the flow path drive unit 29 are provided with a drive seal 74 in order to prevent corrosion and waterproofing of the drive mechanism during sterilization. Inside the automatic culture apparatus 10, there is a groove 45 for installing each base 18. When the base 18 is installed using these grooves 45, the driving units 19 and the base 18 are connected to each other, and a driving force is brought to the base 18 to control the flow path 27.

  The automatic culture apparatus of the present embodiment has the overall configuration as described above. Hereinafter, main components of the automatic culture apparatus of Example 1 will be described.

<Base configuration>
The configuration of the base 18 will be described with reference to FIGS. 4A to 4D, FIGS. 5A to 5D, and FIGS. 6A to 6E.

  4A and 4B are schematic views showing the configuration of the flow path 27 and the base 18, and FIGS. 4C and 4D are views showing the structure of the seal between the cell culture chamber 13 and the refrigerator 14. The individual elements in FIGS. 4A to 4D will be described sequentially.

  4A shows a state in which the channel 27 shown on the left side and the base 18 shown on the right side are separated, and FIG. 4B shows the channel-equipped base 51 in which the channel 27 and the base 18 are integrated. ing. 4A and 4B, 46 shows a seal for sealing the cell culture chamber 13 and the refrigerator 14, and FIG. 4C shows the seal 46 to the claw portion 36-2 provided on the wall of the cell culture chamber 13 and the refrigerator 14. The structure before installation is shown, and FIG. 4D shows the installation state of the seal 46 on the claw portion 36-2. The seal 46 is provided with a hole 47 and a notch 48.

  5A to 5D are schematic views showing a base set used for installing the base 18 in the automatic culture apparatus, its transporter, and a process of connecting the base 18 to the base set 50. FIG. The insertion mechanism 61 of the base set 50 in the figure includes a lever 52, a claw 53, a slide 60, and a clamp 62.

  FIG. 5A shows the state of the base set 50 alone, and FIG. 5B shows the state where the base 51 with a flow path integrated with the flow path 27 is connected to the base set 50. On the other hand, FIG. 5C shows a state in the middle of setting the channel-equipped base 51 to the drive unit 19. FIG. 5D shows the configuration of the transport tool 90 that transports the base set 50.

  6A to 6E are side views regarding the installation of the base 18 using the base set 50, FIG. 6A shows a state where the base 18 is not connected to the base set 50, and FIG. 6B shows that the base 18 is connected to the base set 50. 6C shows a state where the base 18 is moved to the drive unit 19 shown in FIG. 3, and FIG. 6D shows a state where the lever 52 is lifted and the claw 53 of the base 18 is removed and inserted into the groove 45. FIG. 6E shows a state of the base set 50 after the base 18 is installed in the drive unit 19 as shown in FIG.

  Subsequently, in this embodiment, a method for installing the base 18 inside the automatic culture apparatus 10 using the base set 50 will be described. Before the cells are cultured, the automatic culture apparatus 10 is in the state of FIG. 2 in which the cell culture chamber door 11 and the refrigerator door 12 are closed in the state of FIG. Then, the inside is sterilized with a sterilizing gas such as hydrogen peroxide or ozone.

  On the other hand, the flow path 27 and the base 18 are separated as shown in FIG. 4A. The flow path 27 is first manufactured aseptically as a unit, and is put in a sterilization bag or the like. The base 18 is configured to be individually sterilized and is sterilized in a sterilization bag or the like. Also, the base set 50 shown in FIG. 5A, which is used as a guide to the drive unit 19 by setting the base 18 when installing, is configured to be individually sterilized in the same manner as the base 18, and this can also be used as a sterilization bag. Put in and sterilize.

  The base 18, the flow path 27, and the base set 50 are removed from the sterilization bag in a highly clean environment such as a clean bench or a cell isolator in a CPC (Cell Processing Center), and are aseptically placed on the base set 50. And a channel 27, that is, a channel-equipped base 51 is installed. The culture vessel 20 is also connected to the flow path 27 in this highly clean environment and the inside thereof is sealed. Further, the medium 54 is connected so that necessary reagents and washing liquids can be supplied into the flow path 27, such as the medium container 55 and the cell suspension 56.

  First, the base 18 is installed in the base set 50. The claw 53 of the base set 50 is caught by the claw receiver 58 of the base 18 to fix the position. The tube of the flow path 27 has a guide 59 (including authentication by color and number) such as a groove shape to be installed in the base 18, and can be definitely installed by connecting along the guide 59.

  In a state where the culture vessel and the flow path are connected, the inside has a sealed structure. Even when the culture vessel and the flow path are connected and the cells, the culture solution, and the washing solution are introduced, the inside has a sealed structure.

  4B shows a state where the flow path 27 is installed in the guide 59 shown in FIG. 4A, and FIG. 5B shows a state where the base 51 with flow path is installed in the base set 50. The flow path 27 has a sealed structure as described above, and in this state, even if the cleanliness of the external environment is lowered, the inside is in a state of maintaining a high clean environment. Therefore, the base set 50 may be taken out from a high clean environment.

  Then, it is moved to the automatic culture apparatus 10 in the state of FIG. 3, the base set 50 is installed, and the base 18 is connected to the drive unit 19 by the groove 45 and the slide 60. Next, a side view of the insertion mechanism 61 including the slide 60, the lever 52, and the claw 53 that brings about the connection will be described.

  First, when the base 18 is not installed in the base set 50 as shown in FIG. 5A, the insertion mechanism 61 is in the state shown in FIG. 6A. The insertion mechanism 61 can move the slide 60 in and out by pushing and pulling the lever 52. The slide 60 has an L-shaped clamp 62, a claw 53 at one end, and a spring 63 at the other end. Normally, the tab 53 is in a protruding state. 5B and 6B show a state where the base 18 is installed in the base set 50. FIG. As shown in FIG. 5B and FIG. 6B, the claw 53 is installed so as to fit the claw receiver 58 of the base 18. At this time, the base 18 is fixed.

  Subsequently, the base set 50 is moved to the automatic culture apparatus 10 in FIG. 3, the slide 60 is installed in the groove 45 of the automatic culture apparatus 10, the lever 52 is pushed, and the slide 60 is taken out as shown in FIG. 6C. When the slide 60 comes out, the claw 53 is removed from the claw receptacle 58 by pushing the lever 52 upward as shown in FIG. 6D. The state at this time is shown in FIG. 5C. Here, the base 18 is connected to the drive unit 19 of FIG. At this time, the base 18 can be fixed to the drive unit 19 using a stopper or the like. Pulling the lever 52 and putting the slide 60 into the state of FIG. 6E while the lever 52 is tilted, return the lever 52 to the state of FIG. 6A and remove the base set 50 from the automatic culture apparatus 10. Depending on the method, it can be left.

  Here, as shown in FIG. 4C, there is a seal 46 that seals the cell culture chamber 13 and the refrigerator compartment 14 in the flow path 27 between the culture medium base 35 and the flow path base 28. The seal 46 has a notch 48 through which the flow path 27 is set in the hole 47. When the seal 46 is set in the claw 36 as shown in FIG. 4D, the notch 48 is closed, and the channel 27 can be passed through the hole 47 while being sealed. Thereafter, the cell culture chamber door 11 and the refrigerator door 12 are closed, and culture is started. When the base 18 is removed, the above method may be executed in reverse.

Here, the carrier 90 of the base set 50 will be described with reference to FIG. 5D. The transport tool 90 is provided with a moving mechanism 91 for moving the base set 50 up and down by turning a handle 93 and a handle 92 for facilitating transport. The base 18 in which the flow path 27 is set, that is, the base 51 with flow path is installed in the base set 50, and the operator moves it to the automatic culture apparatus 10 to easily set the base 18 to the drive unit 19 Can be.
<Connection between base and mechanism>
A method for driving the valve of the flow path on the base 18 will be described with reference to FIGS. 7A to 7F.

  7A shows the state of the base 18 and the drive unit 19 where the flow path 27 is not connected to the valve 24, and FIG. 7B shows the state where the flow path 27 is connected to the valve 24 of the base 18 and the base 18 and drive unit 19 are connected. FIG. 7C shows a state in which the base 18 and the drive unit 19 are connected as shown in FIG. Further, FIG. 7D shows a state in which a driving force is applied from the driving unit 19 and the valve 24 of the base 18 is driven. In this configuration, a configuration using a motor 69 for applying a driving force is adopted.

  The configuration and operation of the valve 24 will be described with reference to FIGS. 7A to 7D. First, the case where the valve 24 can be separated using the motor 69 will be described. The base 18 includes a valve mechanism 65, a spring 66, and a switching mechanism 67. These are sterilizable materials. One tube 68 of the flow path 27 is inserted between each of the valve mechanism 65 and the switching mechanism 67, and one of the tubes 68 is pushed above and below the switching mechanism 67 to close one of the tubes.

  FIG. 7A shows the state before the tube 68 is inserted, and FIG. 7B shows the state after the tube 68 is inserted. Normally, the switching mechanism 67 is pushed by the spring 66, and the lower tube 68 is pushed as shown in FIG. 7B. The drive unit 19 includes a motor 69, an encoder 70, a rod 71, and a cam 72. The cam 72 is protected by an elastic drive seal 74 for protection. Since the motor 69, the encoder 70, the wiring 79, etc. cannot withstand sterilization processing, the inside of the drive unit 19 and the outside are isolated, and the internal electronic components are not affected even if the outside is sterilized. As shown in FIG. 7B, normally, before the base 18 and the drive unit 19 are connected, the cam 72 does not come out of the upper surface of the drive unit 19. Reference numeral 74 denotes a drive seal made of an elastic body.

  And the state which connected the base 18 to the drive part 19 as mentioned above is shown to FIG. 7C. The cam 72 comes to a position at the lower end of the switching mechanism 67. Here, when the motor 70 is rotated by a predetermined amount while being measured by the encoder 71, the cam 72 pushes up the switching mechanism 67, the valve mechanism 65, the tube 68 that has been closed until then opens, and the tube 68 that has been opened closes. A path switch is performed. The state is shown in FIG. 7D. The drive seal 74 extends as the cam 72 moves up.

  By rotating the motor 69 to the original state in this state, the initial state shown in FIG. 7C can be restored, and the base 18 can be removed as shown in FIG. 7B.

  In the above configuration, the motor 69 is used for switching the tube of the base. However, as a modification thereof, a solenoid can be used instead of using the motor.

  7E and 7F, 75 is a solenoid, 76 is a rod, 77 is a spring, 78 is a connector, and 79 is a wiring. FIG. 7E shows a state in which the solenoid 75 of the valve 24 is separated, and FIG. 7F shows a state in which the solenoid 68 is connected and the tube 68 is switched.

  7A, the rod 76 is pulled by the spring 77 as shown in FIG. 7E. When the base 18 is connected to the drive unit 19 and the solenoid 75 is actuated, a state corresponding to FIG. 7D is obtained, and the rod 76 pushes out the spring 66, and the tube 68 is switched as shown in FIG. 7F. The solenoid 75 used here has the same function as a push solenoid.

<Injection of cell suspension or medium into culture vessel>
8A to 8D, a method for injecting cells, that is, the cell suspension 56, or the culture solution, that is, the culture medium 54 into the culture container 20 by the automatic culture apparatus 10 will be described. 8A shows a state before the base 18 is connected, FIG. 8B shows a state after the base 18 is connected, FIG. 8C shows a state where the culture vessel base 21 is set up vertically, and FIG. 8D shows a culture vessel base. 21 shows a state where 21 is vertically lowered.

  As for the inside of the automatic culture apparatus 10, the flow path 27 and the base 18 are not set to the drive part 19 like FIG. 8A after internal sterilization. When the base 18 in which the flow path 27 is set in the base set 50 shown in FIG. 5 is attached to the inside of the automatic culture apparatus 10 using the seal 46, the result is as shown in FIG. 8B. In this state, the cells in the culture vessel 20 are usually cultured. The inside of the refrigerator 14 that is a heat retaining unit maintains approximately 4 ° C. The cell culture chamber 13 includes a heater 39, a carbon dioxide supply mechanism 30, a humidity generation mechanism 73, and respective sensors for adjusting the carbon dioxide concentration of 5% and humidity of 100% to 37 ° C. Then, the internal air is circulated to keep the environment inside the cell culture chamber 13 uniform. If necessary, a HEPA filter or the like can be bitten around the fan 38 to keep the inside of the cell culture chamber 13 clean. In order to maintain this environment, the internal environment is monitored, adjusted, and controlled by the control unit 15 and the display unit 42 and the input unit 43 installed on the wall of the casing in which the control unit 15 is built. Further, the state of the cells can be grasped by an inspection mechanism that is separately prepared in the automatic culture apparatus 15 and includes a microscope, a medium sensor, etc. (not shown).

  Next, a method for supplying the cell suspension 56 and the culture medium 54 to the culture vessel 20 will be described. First, the motor 23 of the rotating mechanism 17 is driven to stand the culture vessel base 21 and the culture vessel drive unit 22 vertically as shown in FIG. 8C. At that time, the cell suspension 56 and the culture medium 54 in the refrigerator 14 control the flow path base 28 and the flow path drive unit 29, that is, the valve 24, the motor 25, and the syringe pump 64. Move to tank 26 and warm to 37 ° C. The warmed cell suspension 56 and the culture medium 54 are placed in a vertical culture container by controlling the flow path 27 by the flow path base 28 and the flow path drive unit 29 and the culture vessel base 21 and the culture container drive unit 22. 20 from below. Thereby, the bubble which has entered the inside of the culture vessel 20 can be excluded from the upper side, and the inside can be efficiently filled with the liquid. When the desired amount of cell suspension 56 and medium 54 are supplied, the motor 23 is driven to cultivate the cells as shown in FIG. 8B. At this time, if the waste liquid in the culture vessel 20 can be collected, the culture solution is moved to a separately prepared inspection mechanism such as a medium sensor.

  Furthermore, one means for efficiently discharging the medium in the culture vessel 20 will be described. The motor 23 is driven to stand the culture vessel base 21 and the culture vessel drive unit 22 vertically in the reverse direction of FIG. 8C as shown in FIG. 8D. And all the waste liquid in the culture container 20 can be discharged | emitted by taking out a waste liquid from the lower side of the culture container 20 of this state. 8C and 8D, the liquid flows in one way from the right to the left in the culture container 20 in the state of FIG. 8B when the liquid in the culture container 20 is injected and discharged. Thus, since the waste liquid does not return to the culture space in the culture vessel 20 and the cells are always supplied with a clean and fresh medium, it is possible to maintain a clean environment without entering bacteria or the like.

<Circuit configuration of automatic culture equipment>
FIG. 9 is a block diagram showing a configuration of a control system circuit in the control unit 15 for controlling internal devices in the automatic culture apparatus 10.

  The control system circuit of the control unit 15 of the automatic culture apparatus 10 includes an input unit (keyboard, mouse, etc.) 81 for inputting data and instructions, and controls each operation of the automatic culture apparatus 10, for example, a central processing unit (Central A control processing unit 82 composed of a processing unit (CPU), a display unit 80 showing the control status to the user, a ROM 85 for storing programs and parameters, a RAM 86 for temporarily storing data and processing results, A memory 85 for performing an operation such as a cache, a sterilization process, a heater, a fan, a carbon dioxide supply, a water supply, and the like are performed, and an environment holding device 87 using a sensor for monitoring these conditions is connected to the base 18. A culture vessel drive unit 22, a flow channel drive unit 29, a refrigerator control 88 for controlling the environment in the refrigerator, and a motor 23 for rotating the culture vessel drive unit 22. That.

  When the user instructs a culture process to be processed from the input unit 81 or the communication unit 84, the control processing unit 82 controls the inside of the automatic culture device 10 with the sterilization function of the environment holding device 87 according to the culture preparation program stored in the ROM 85. Sterilization is performed, and after the treatment is completed, the culture environment holding treatment is advanced to a temperature of 37 ° C., a 5% carbon dioxide concentration, a humidity of 100%, and a clean environment field. At the same time, the refrigerator control 88 is performed. When the control processing unit 82 detects the set of the base 18 with the position sensor according to the automatic culture program stored in the ROM 85, the control processing unit 82 includes the culture vessel driving unit 22, the flow path driving unit 29, and the motor 23 of the rotating shaft mechanism 17. A cell culture process in the culture vessel 20 is performed by the drive unit 19. At any time, the display unit 80 and the communication unit 84 can indicate the processing status to the user. When the cell culture process is completed, the display unit 80 and the communication unit 84 indicate the completion to the user, and when the removal of the base 18 is detected, the control processing unit 82 performs the termination process according to the termination program stored in the ROM 85. As described above, a series of cell culture processes by the automatic culture apparatus 10 can be realized.

  Although the first embodiment has been described in detail above, the automatic culture apparatus of the second embodiment having a different base configuration will be described with reference to FIGS. 10A to 10D. In the description, only differences from the first embodiment will be described, and description of the same parts as those of the first embodiment will be omitted. In this embodiment, the configuration of an automatic culture apparatus that uses a base that does not separate valves will be described.

  10A and 10B, a method of installing the base 18 'in the case of the valve 24 that is not separated will be described. FIG. 10A shows a state where the base 18 ′ is installed in the automatic culture apparatus 10, and FIG. 10B shows a case where the base 18 ′ is removed.

  Compared to FIGS. 1 and 2, the valve 24 and the motor 25 are included in the culture vessel base 21 ′ and the flow path base 28 ′ of the base 18 ′, and therefore there is no drive unit 19 as shown in FIG. In addition, a culture vessel base pedestal 94 and a flow path base pedestal 95 having a groove 45 are provided. Further, there is no drive seal 74 for protecting the cam 72 and rod 76 of the valve 24. However, since the base 18 'can be separated, a wiring 79 for the valve 24 and the motor 25 and a connector 78 for connecting the same are necessary. The connector 78 is connected by the carrier 90 and the base set 50 when the base 18 is installed.

  FIGS. 10C and 10D respectively correspond to FIGS. 4A and 4B of the first embodiment, and show a configuration in which a valve 24 and a motor 25 constituting a drive unit are included in the base 18 ′. FIG. 10C shows a state where the flow path 27 and the base 18 ′ are separated, and FIG. 10D shows a state where the flow path 27 and the base 18 ′ are integrated.

  According to the present embodiment, since the drive unit 19 having the shape shown in FIG. 1 is not necessary for the automatic culture apparatus 10, precise positioning accuracy is not required at the time of installation, and it can be installed more easily. It is. However, since wiring for the valve 24 and the motor 25 is necessary at the time of installation, it is necessary to connect the connector 78, and it is desirable that the connector 78 has waterproof and gas-proof performance.

  In addition, according to the present embodiment, since the drive unit is included in the base 18 ′, they can be attached to and detached from the automatic culture apparatus together, no drive seal is required, and sterilization is easy. That is, even if the sterilization process or the decontamination process is performed inside the automatic culture apparatus in a state where the culture container 20, the flow path 27, and the base 18 'are removed, the drive unit is not damaged.

  Furthermore, as a modification of the present embodiment, a case where the solenoid 75 is also included in the base 18 ′ including the valve 24 will be described with reference to FIGS. 11A and 11B.

  FIG. 11A shows a configuration when the solenoid 75 is not operating, and FIG. 11B shows a state where the solenoid 68 is operated and the tube 68 is switched. The contents described here mainly relate to the valve 24, and the drive system such as the syringe pump 64 on the base 18 'shown in FIG. 1 has the same configuration as the operation of FIGS. 7A to 7D.

  In this configuration, in the same state as FIG. 7A, the rod 76 is pushed out by the spring 77 as shown in FIG. 11A. Here, the base 18 ′ has a connector 78 for connecting to the wiring 79. This connector 78 is desirably waterproof and gas-proof. When the solenoid 75 is operated, the rod 76 is pulled by the solenoid as shown in FIG. 11B, and the tube 68 is switched. The solenoid 75 used here has the same function as the pull solenoid.

  As mentioned above, although several embodiment of this invention was described, it cannot be overemphasized that these embodiment is only an example for implement | achieving this invention, and does not limit the technical scope of this invention.

  INDUSTRIAL APPLICABILITY The present invention is useful as an automatic culture apparatus for culturing cells, particularly as a system for automatically culturing by supplying a driving force with a culture vessel and a flow path having a sealed structure.

DESCRIPTION OF SYMBOLS 10 ... Automatic culture apparatus 11 ... Cell culture room door 12 ... Refrigerator door 13 ... Cell culture room 14 ... Refrigerator 15 ... Control part 16 ... Clean air circulation part 17 ... Rotating mechanism 18, 18 '... Base 19 ... Drive part 20 ... Culture Container 21, 21 '... Culture vessel base 22 ... Culture vessel drive unit 23 ... Motor 24 ... Valve 25 ... Motor 26 ... Tank 27 ... Channel 28, 28' ... Channel base 29 ... Channel drive unit 30 ... Carbon dioxide supply Mechanism 31 ... Carbon dioxide sensor 32 ... Temperature sensor 33 ... Humidity sensor 34 ... Fan 35 ... Medium base 36 ... Claw part 37 ... Exhaust port 38 ... Fan 39 ... Heater 40 ... Sterilization gas supply port 41 ... Sterilization gas discharge port 42 ... Display Part 43 ... Input part 44 ... Communication part 45 ... Groove 46 ... Seal 47 ... Hole 48 ... Cut 50 ... Base set 51 ... Base 52 with flow path ... Lever 53 ... Claw 54 ... Medium 55 ... Medium container 56 Cell suspension 57 ... Cell suspension container 58 ... Claw receptacle 59 ... Guide 60 ... Slide 61 ... Insertion mechanism 62 ... Clamp 63 ... Spring 64 ... Syringe pump 65 ... Valve mechanism 66 ... Spring 67 ... Switching mechanism 68 ... Tube 69 ... motor 70 ... encoder 71 ... rod 72 ... cam 73 ... humidity generating mechanism 74 ... drive seal 75 ... solenoid 76 ... rod 77 ... spring 78 ... connector 79 ... wiring 80 ... display part 81 ... input part 82 ... control processing part 83 ... Memory 84 ... communication unit 85 ... ROM
86 ... RAM
87 ... Environment holding device 88 ... Refrigerator control 90 ... Transport tool 91 ... Vertical mechanism 92 ... Handle 93 ... Handle 94 ... Culture vessel base pedestal 95 ... Channel base pedestal.

Claims (5)

An automatic culture apparatus capable of supplying a culture medium from a culture medium supply means in the first chamber to a culture container provided in the second chamber via a flow tube,
The first chamber and the second chamber have a recess in the opposing wall surface,
A seal member that is detachable from the recess,
The automatic culture apparatus, wherein the seal member is provided with a hole through which the flow channel tube can pass. The automatic culture apparatus according to claim 1,
An automatic culture apparatus, wherein a space is provided between the first chamber and the second chamber. The automatic culture apparatus according to claim 1 or 2,
The first chamber is a refrigerator;
The automatic culture apparatus, wherein the second chamber is a culture chamber. An automatic culture apparatus having a refrigerator room and a culture room,
In the refrigerator compartment, a culture medium base in which a culture tank is installed is detachably installed,
In the culture chamber, a channel base on which a channel tube is installed is detachably installed,
The wall having the refrigerator compartment and the culture chamber adjacent or in contact with each other has a recess,
A seal member that is detachable from the recess,
The seal member is provided with a hole,
The automatic culture apparatus, wherein the flow path tube installed on the flow path base is connected to the medium tank in the refrigerator compartment through the hole. The automatic culture apparatus according to claim 4,
An automatic culture apparatus, wherein a space is provided between the culture chamber and the refrigerator compartment.

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