JP2005027607A - Automatic culture apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To prevent the leakage of atmosphere from a culture chamber to the outer environment during a long-running culture process. <P>SOLUTION: The automatic culture apparatus 1 is provided with a 1st space S1 containing a culture chamber 4 removably holding a culture vessel 3 containing cells and culturing the cells while keeping a prescribed culture condition, a 2nd space S2 holding a treating part 32 to apply a prescribed treatment to the cells in the culture vessel 3, and a transfer mechanism 9 to transfer the culture vessel 3 through a connection port 33 placed between the 1st space S1 and the 2nd space S2. The connection port 33 is closed with an openable door 2 and the air near the connection port 33 is sucked with a suction means when the openable door 2 is opened. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to an automatic culture apparatus.

As a conventional automatic culture apparatus, an apparatus including a fixed storage shelf that can store a plurality of culture vessels and a conveying means that can move horizontally, vertically, and rotationally is known (see, for example, Patent Document 1). .)
This automatic culture apparatus includes a plurality of small rooms arranged in a vertical direction on a storage shelf arranged in a culture chamber, and accommodates one culture container in each small room for culturing, either in the middle of culture or in culture At the end, the conveying means is operated to take out the culture containers one by one from the small room, or to accommodate the culture containers in the small room.
In this automatic culture apparatus, each small chamber is opened in the culture chamber without being sealed, and the culture chamber is maintained at a constant culture condition.

Moreover, in this automatic culture apparatus, the culture container taken out from the small room is sent out of the culture room through an openable and closable opening provided on the wall surface of the culture room, and the medium is obtained by various processing devices arranged outside the culture room. Processing such as replacement is performed.
JP 2002-262856 A (FIG. 1 etc.)

  However, in such a conventional automatic culture apparatus, the opening of the culture chamber is opened each time the culture container is taken out of the culture chamber. In this case, the culture chamber is maintained in an atmosphere that satisfies a predetermined culture condition, and there is a disadvantage that the atmosphere in the culture chamber leaks out of the culture chamber each time the opening is opened. In particular, since the culture chamber is maintained at a high temperature and high humidity, if high humidity air leaks to the outside, there may be a disadvantage that various processing apparatuses disposed outside are adversely affected. In addition, the higher the humidity, the higher the possibility of microbial infection in the cell treatment apparatus.

  This invention is made | formed in view of the situation mentioned above, Comprising: It aims at providing the automatic culture apparatus which can prevent that the atmosphere in a culture chamber leaks out of a culture chamber in the culture | cultivation process extended over a long period of time.

In order to achieve the above object, the present invention provides the following means.
The invention according to claim 1 accommodates the culture vessel containing the cells in a removable manner, and accommodates the first space for accommodating the culture chamber for culturing the cells while maintaining the predetermined culture conditions, and the culture vessel is accommodated in the culture vessel. A second space in which a processing unit for performing a predetermined process on the cells is disposed; and a transport mechanism for transporting the culture vessel through a communication port provided between the first space and the second space. An automatic culture apparatus provided with an opening / closing door for opening and closing the communication opening at the communication opening, and an intake means for sucking air around the communication opening when the opening / closing door is opened. provide.

  According to the present invention, the cells cultured in the culture chamber of the first space are taken out from the culture chamber together with the culture container, and are transported into the second space through the communication port by the operation of the transport mechanism. Then, after a predetermined process such as medium replacement is performed in the processing unit arranged in the second space, the transport mechanism is operated again to be transported into the first space through the communication port, and the culture is performed in the culture chamber. Will continue.

  In this case, when the culture vessel is not passed through the communication port, the communication port is closed by operating the open / close door, the first space and the second space are sealed, and the air in both spaces is mutually connected. It is held so as not to be mixed in. On the other hand, when the culture vessel is allowed to pass through the communication port, the communication port is opened by operating the open / close door, but even in this case, the air around the communication port is sucked by the operation of the intake means, The air in both spaces is held so as not to mix with each other. The humidity in the second space of the cell treatment device is kept low because the atmosphere in the first space containing the culture chamber is unlikely to flow out into the second space, and the air in the second space of the cell treatment chamber keeps the culture chamber. Since the second space of the cell processing chamber is hardly contaminated with microorganisms, the contamination does not easily expand because it is difficult to flow out to the accommodated first space.

According to a second aspect of the present invention, there is provided the automatic culture apparatus according to the first aspect, wherein the automatic culture apparatus includes a control device that operates the suction means before the opening / closing door is opened.
According to the present invention, since the opening / closing door is opened after the intake means is operated by the operation of the control device, when the opening / closing door is opened, the suction by the intake means has already started, and the air in both spaces Can be efficiently prevented from flowing into another space through the communication port.

According to a third aspect of the present invention, in the automatic culture apparatus according to the first or second aspect, the intake means includes an intake port on at least one of the first space side and the second space side of the communication port. An automatic culture apparatus is provided.
According to the present invention, when the air inlet is provided on the first space side, the air in the first space is sucked into the air inlet by the operation of the air intake means, and the first air via the communication port. The air in the second space that has entered the space is also sucked into the intake port. Therefore, the inflow of air is prevented in the vicinity of the communication port, and both spaces can be maintained in different atmospheres.

  According to the automatic culture apparatus according to the present invention, the atmosphere in the first space containing the culture chamber is prevented from flowing out to the second space through the communication port, and the process arranged in the second space. The atmosphere of the part can be suitably maintained. Moreover, the atmosphere in the second space can be prevented from flowing into the first space. Since the atmosphere in each space is not easily mixed, a steady atmosphere with low humidity is maintained, and even when the atmosphere is contaminated, it is possible to prevent the atmosphere from easily flowing out to other spaces.

An automatic culture apparatus according to an embodiment of the present invention will be described below with reference to FIGS.
As shown in FIG. 1, the automatic culture apparatus 1 according to the present embodiment is sealed with a transparent wall material that can be observed from the outside, and communicates with each other via a shutter (opening / closing door) 2. And a second space S2.

  A total of four culture apparatuses 4 for accommodating the culture vessels 3 are arranged in each of the two side spaces S11 and S13 of the first space S1, and a transport robot (transport) for moving the culture vessels 3 is placed in the central space S12. Mechanism) 5 is provided. In the upper part of the central space S12, an air purifier 6 for sending a clean descending air flow for purifying the air in the central space S12 is provided.

The four culture apparatuses 4 are arranged with a space between the two culture apparatuses 4 arranged side by side with the doors 4a facing each other by arranging the doors 4a toward the central space S12.
As shown in FIGS. 2 and 3, each culture device 4 includes an opening 4b on one side surface and includes a door 4a that can open and close the opening 4b. A plurality of rail-like tray holding members 4c are provided at corresponding height positions on the left and right side walls toward the opening 4b. 7 can be accommodated in a plurality of stages in the vertical direction. Each culture apparatus 4 is maintained at predetermined culture conditions such as a temperature of 37 ± 0.5 ° C., a humidity of 100%, and a CO ₂ concentration of 5%. The tray holding member is not limited to a rail shape, and may be in any form as long as it can support the tray so that it can be taken in and out.

  A plurality of, for example, ten culture vessels 3 can be placed side by side on each tray 7. As shown in FIG. 4, each culture vessel 3 includes a vessel body 3a and a lid 3b provided on the upper surface of the vessel body 3a. A protrusion 3c that is hooked by a hand in the space S2 is provided.

  Below each culture apparatus 4 is disposed a stocker 8 that accommodates a plurality of unused culture containers 3 mounted on a tray 7. The stocker 8 has a door 8a that can be opened and closed on the side facing the outside of the first space S1 on the side opposite to the door 4a of the culture apparatus 4. The door 8a is formed in a size that opens one entire side surface of the stocker 8.

  The transfer robot 5 is arranged at approximately the center of the interval between the four culture apparatuses 4. The transfer robot 5 includes a first arm 5a that can rotate horizontally, a second arm 5b that is connected to the tip of the first arm 5a so as to be rotatable about a vertical axis, and a vertical axis that is connected to the tip of the second arm 5b. A hand 5c that is rotatably mounted around itself and does not have a mechanism for deteriorating the environment in the culture chamber such as a drive unit and a conduction mechanism, and the first arm 5a, the second arm 5b, and the hand 5c can be moved up and down. And an elevating mechanism 5d. As a result, the transfer robot 5 accesses all the culture vessels 3 in the four culture apparatuses 4 and also crosses the shutter 2 and is disposed between the first space S1 and the second space S2. It has a horizontal operating range in which the culture vessel 3 can be handed over.

The conveyor 9 includes two endless belts 9a arranged on the left and right sides with a gap larger than the width dimension of the hand 5c of the transport robot 5, so that the tray 7 can be placed over the endless belts 9a. It has become. Further, the transfer robot 5 has access to all the trays 7 in the culture apparatus 4 and has a vertical operation range in which at least the uppermost tray 7 in the stocker 8 can be accessed.
The belt 9a is not limited to an endless belt.

  The hand 5c is formed in a flat shape extending in the horizontal direction so that the tray 7 can be placed, and has a thickness dimension that can be inserted into a gap between the trays 7 accommodated in the culture apparatus 4. . The hand 5c is lifted from the state inserted in the gap between the trays 7, so that the tray 7 is pushed up from below by the two arms and taken up from the tray holding member 4c, and the tray 7 is stably held. It can be done.

  The second space S2 is formed in a transparent housing 30 that is visible from the outside. A processing unit 32 is provided in the second space S2, and is divided into an upper space S21 and a lower space S22 by a horizontal partition wall 31 as shown in FIGS. Yes.

In the second space S2, a handling robot 10 that handles the culture vessel 3 conveyed from the first space S1 by the conveyor 9 with the shutter 2 opened, and a centrifuge for separating cells from the culture medium in the culture vessel 3 A separator 11, two dispensing robots 13 that are equipped with an electric pipette 12 for dispensing various liquids such as serum and reagents, and that can be horizontally rotated and moved up and down, and an electric pipette 12 for these dispensing robots 13. A plurality of disposable chips 14 to be attached to the tip are accommodated and three chip supply devices 15 that can be provided within the operating range of the dispensing robot 13 and various liquids such as serum and reagents are stored in a plurality of containers. A reagent supply device 16, a microscope 17 capable of observing the state of cells in the culture vessel 3, and a waste liquid discarded due to exchange of each reagent and culture medium are respectively stored. A number of storage tanks 18, a horizontal movement mechanism 19 that moves the culture container 3 so that the culture container 3 can be delivered between the conveyor 9 and the robots 10 and 13, and a slider 20 of the horizontal movement mechanism 19. And a mounting table 21 on which the received culture vessel 3 is mounted.
The second space S2 is also provided with an air purifier (not shown) that sends a clean downward airflow to purify the air in the second space S2.

  The handling robot 10 is a horizontal articulated robot that horizontally moves and raises and lowers a gripping hand 25 a that handles the culture vessel 3. For example, the example shown in FIG. 1 includes three horizontal arms 10b, 10c, and 10d connected to each other, and an elevating mechanism 10e that moves the horizontal arms 10b to 10d up and down. In addition to the gripping hand 10a for gripping the culture vessel 3 at the tip of the horizontal arms 10b to 10d, an electric pipette (not shown) that can attach and detach a chip 14 for taking in and out cells and culture medium from the culture vessel 3, A lid opening / closing hand (not shown) for opening and closing the lid 3b of the culture vessel 3 is provided.

  The handling robot 10 opens and closes the lid 3b of the culture vessel 3 on the tray 7 that has been conveyed by the conveyor 9, and grips and conveys the culture vessel 3 to supply it to the mounting table 21 and the microscope 17, and the electric pipette The tip tip 14 is replaced, and the cell-containing medium taken out from the culture vessel 3 is put into the centrifuge 11. Therefore, the handling robot 10 arranges various devices such as the conveyor 9, the mounting table 21, the microscope 17, the chip supply device 15, the chip collection device (not shown), and the centrifuge 11 within the operation range.

  The centrifuge 11 is configured to separate and sink the cells with heavy specific gravity suspended in the medium by rotating the medium containing the cells supplied from the handling robot 10 at a low speed.

  The dispensing robot 13 includes a horizontally rotatable arm 13a having an electric pipette 28 to which the tip 14 is detachably attached at the tip, and an elevating mechanism 28b for raising and lowering the arm 13a. The dispensing robot 13 supplies a culture medium and various reagents into the culture vessel 3 that has been transported by the horizontal movement mechanism 19. Therefore, the dispensing robot 13 arranges various devices such as the mounting table 21 on the horizontal movement mechanism 19, the chip supply device 15, the chip collection device and the reagent supply device 16 within the operation range.

  The tip supply device 15 accommodates a plurality of tips 14 in an arrayed state in a container 15a opened upward with the attachment port to the electric pipette 12 facing upward, and the handling robot 10 and the dispensing robot 13 are When a new tip 14 is required, the tip 14 is attached to the tip of the electric pipette 12 simply by inserting the electric pipette 12 from above. The container 15a is attached to the moving mechanism 10c so as to be reciprocally moved between the operating range of the handling robot 10 and the dispensing robot 13 and the lid 15b, and when the tip 14 need not be replaced, the tip 14 In order to prevent adhesion of dust or the like to the head, the moving mechanism 15c is operated to be disposed below the lid body 15b.

  The tip collection device includes a gripping device that grips the tip 14 at the inlet of the collection container. When the tip 14 used in the handling robot 10 or the dispensing robot 13 is inserted into the gripping device, the tip collection device grips the tip 14. It is supposed to be. In this state, the handling robot 10 and the dispensing robot 13 move the electric pipette 12 to remove the used tip 14 from the tip of the electric pipette 12 and collect it in the collection container.

  For example, as shown in FIG. 3, the reagent supply device 16 accommodates a horizontally rotatable table 16a inside a cylindrical casing, and a cylindrical shape having a fan-shaped bottom shape on the table 16a. A plurality of the reagent containers 16b are arranged and mounted in the circumferential direction. Various reagents etc. are stored in each reagent container 16b. For example, serum such as MEM (Minimal Essential Medium), DMEM (Dulbecco's Modified Eagle Medium), FBS (Fetal Bovine Serum) or human serum, which constitutes a medium necessary for culturing cells Proteolytic enzymes such as trypsin that exfoliate cells in the culture vessel 3, growth factors such as cytokines that grow cells during culture, differentiation inducers such as dexamethasone that differentiates cells, and penicillin antibiotics Antibiotics, hormones such as estrogens, and nutrients such as vitamins are stored.

  An insertion port 16 c through which the dispensing robot 13 inserts the tip 14 at the tip of the electric pipette 12 is provided on the upper surface of the casing of the reagent supply device 16. The insertion port 16 c is disposed within the operation range of the dispensing robot 13. Each reagent container 16b includes an opening (not shown) disposed on the upper surface thereof at a position corresponding to the insertion port 16c. Thereby, the dispensing robot 10 disposes the tip 14 at the tip of the electric pipette 12 from above by placing the opening of the reagent container 16b vertically below the insertion port 16c of the casing by rotating the table 16a. The reagent etc. which are inserted in 16b and are stored inside can be aspirated. Two reagent supply devices 16 and two dispensing robots 13 are provided so that a chemical solution such as trypsin common to the sample and a liquid such as serum inherent to the sample are separated and handled. This is because.

  The microscope 17 is used in the case of observing the state of cells in the culture vessel 3 and the degree of proliferation or counting the number of cells in the middle of the culture process or at the time of medium exchange. Yes. The XY stage, working distance adjustment, magnification change, and the like of the microscope 17 are all configured to be performed by remote control. By disposing the eyepiece lens toward the outside of the second space S2, the state of the cells in the culture vessel 3 may be viewed from the outside of the automatic culture apparatus 1.

  The storage tank 18 stores, for example, DMEM or PBS (phosphate buffered saline) that can be used in common for all specimens, and a reagent container 16a in the reagent supply apparatus 16 as necessary. It is designed to supply inside. In addition, the storage tank 18 may be a waste liquid tank that stores waste medium and the like that are discharged during medium replacement.

  The horizontal movement mechanism 19 includes a slider 20 that can be moved in the horizontal direction by a linear movement mechanism. The mounting table 21 is mounted on the slider 20, and the culture vessel 3 mounted on the mounting table 21 can be moved from the conveyor 9 to the operating range of the dispensing robot 13.

The mounting table 21 includes a holding mechanism (not shown) for mounting and holding the culture vessel 3 transferred from the tray 7 on the conveyor 9. Further, the mounting table 21 may include a vibration device (not shown) that applies vibration to the culture vessel 3 that is mounted. For example, in addition to a device that reciprocally swings the culture vessel 3 within a predetermined angle range, a device that applies ultrasonic vibration or a device that applies horizontal vibration may be employed as the vibration device.
A control device (not shown) is connected to various devices of the automatic culture device 1 according to the present embodiment. The control device controls the order of each process, operation timing, and the like, and records and saves operation history and the like.

  The first space S1 and the second space S2 communicate with each other through a communication port 33 as shown in FIGS. 1 and 5 to 7. The communication port 33 can be opened and closed by the shutter 2. The communication port 33 is provided with the conveyor 9 on both sides of the lower portion thereof, and an intake port 34 is provided on both the first space S1 side and the second space S2 side of the lower central portion. The intake port 34 passes through a partition wall 31 that vertically partitions the second space S2 and is connected to a suction pump (not shown) disposed in the lower space S22. The shutter 2 and the suction pump are connected to the control device, and the control device operates the suction pump when the communication port 33 is opened by opening the shutter 2, and the first space S1 and the second space Air in the upper space S21 of the space S2 is sucked.

The operation of the automatic culture apparatus 1 according to the present embodiment configured as described above will be described below.
In order to culture bone marrow mesenchymal stem cells using the automatic culture apparatus 1 according to this embodiment, first, the centrifuge 11 in a state where bone marrow fluid collected from a patient is placed in a centrifuge container (not shown). In This step may be performed by an operator or may be performed by the handling robot 10. Thereby, the operation of the centrifuge 11 collects bone marrow cells having a high specific gravity from the bone marrow fluid.

  The collected bone marrow cells are put into the culture vessel 3 by the handling robot 10. At this time, ten empty culture vessels 3 placed on the tray 7 are pushed out from the first space S1 to the second space S2 by the operation of the conveyor 9. The handling robot 10 is transferred onto the mounting table 21 by opening the two lids 3b of the culture vessel 3 thus pushed out and then operating the gripping hand 25a to grip it. A robot that opens the lid 3b may be provided separately. As a result, the lid 3b can be opened immediately before processing, and the probability that foreign matter enters the container body 3a can be reduced.

When the chip supply device 15 operates the moving mechanism 30c to place an unused chip 14 within the operating range of the handling robot 10, the handling robot 10 receives the unused chip 14 from the chip supply device 15 and is electrically driven. Attach to the tip of the pipette 12.
In this state, the handling robot 10 is operated to bring the tip 14 at the tip of the electric pipette 12 into contact with the bone marrow cell suspension submerged in the centrifuge 11. Then, the bone marrow cells are sucked into the tip 14 by operating the electric pipette. The aspirated bone marrow cells are loaded into the culture vessel 3 which is transferred by opening the lid 3b on the mounting table 21 by operating the handling robot 10.

  When the bone marrow cells have been put into the culture container 3, the handling robot 10 carries the chip 14 to the chip collection device and removes the used chip 14. Moreover, the chip supply apparatus 15 arrange | positions the container 15a under the cover body 15b by the action | operation of the moving mechanism 15c.

  Next, the culture vessel 3 into which the bone marrow cells have been placed is moved horizontally along with the mounting table 21 by operating the horizontal movement mechanism 19, and is placed within the operating range of each dispensing robot 13. The dispensing robot 13 operates the electric pipette 12 with the unused tip 14 received from the tip supply device 15 attached to the tip thereof, thereby allowing DMEM, serum, or various kinds of substances from the reagent container 16b of the reagent supply device 16. After a suitable amount of reagent is aspirated, the reagent is conveyed to above the culture vessel 3 and injected into the culture vessel 3. Serum and each reagent are aspirated by exchanging the chip supply device 15 with an unused chip 14 for each aspiration of each reagent and the like. Thereby, in the culture container 3, it exists in the state with which the bone marrow cell was mixed in the appropriate culture medium. In addition, in order to distribute bone marrow cells uniformly in the culture medium, the mounting table 21 may be operated and the whole culture vessel 3 may be vibrated. The culture vessel 3 that has completed all the processing is returned to the operating range of the handling robot 10 by the operation of the horizontal movement mechanism 19. The handling robot 10 covers the culture vessel main body 3a with the lid 3b.

  After predetermined processing is performed on all the culture containers 3 on the tray 7, the culture container 3 placed on the tray 7 is moved from the second space S2 to the center of the first space S1 by operating the conveyor 9. It is inserted into the space S12. In this state, by operating the transfer robot 5, the tray 7 is lifted by the hand 5c. Then, when the tray 7 is transported to the front of the culture apparatus 4 to be accommodated, the tray 7 is inserted on the empty tray holding member 4c. Then, by closing the door 4a again, cells are cultured while maintaining the culture conditions in the culture device 4 constant. Needless to say, the order of input of bone marrow cells, input of DMEM, serum, various reagents, and aspiration may be changed as appropriate.

  Further, when exchanging the medium or the container, the culture container 3 in the culture apparatus 4 is taken out together with the tray 7 by the operation of the transfer robot 5 arranged outside the culture apparatus 4 in the same manner as described above. Passed from the first space S1 to the second space S2. In the second space S2, trypsin is injected into the culture vessel 3 and the cells in the culture vessel 3 are peeled off, and are then introduced into the centrifuge 11 by the operation of the handling robot 10 to be mesenchymal stem cells. Only necessary items are collected. Other processing steps are the same as described above.

  And a mesenchymal stem cell is proliferated to sufficient cell number by performing the culture | cultivation process over a predetermined period through several times of culture medium exchanges and container exchanges. Whether or not the number of cells has reached a sufficient level is observed or measured by transporting the culture vessel 3 with mesenchymal stem cells attached to the bottom surface to the microscope 17 by the operation of the handling robot 10, and the degree of cell proliferation Is judged. In addition, the culture container 3 of the same sample may be mounted on the tray 7, or the culture containers 3 of different samples may be mixed. In addition, the culture container 3 of the same specimen may be placed on the mounting table 21, or the culture containers 3 of different specimens may be mixed.

  In this manner, the automatic culture apparatus 1 according to the present embodiment can automatically cultivate mesenchymal stem cells having a sufficient number of cells from bone marrow fluid collected from a patient. In addition, after sufficient mesenchymal stem cells are obtained, a biological tissue supplement such as calcium phosphate and a differentiation-inducing factor such as dexamethasone are introduced into the culture vessel 3, and the culture process is continued again. You may decide to manufacture the biological tissue filling body which can be compensated for the defect | deletion part of a biological body.

  In this case, according to the automatic culture apparatus 1 according to the present embodiment, in the first space S1, every time the door 4a of the culture apparatus 4 whose interior is maintained in a high temperature and high humidity culture environment is opened, the high temperature and high humidity is obtained. Since the atmosphere is released into the first space S1, the humidity is relatively high. On the other hand, in the second space S2, various mechanism units, a microscope, and the like are arranged. For this reason, in the state where the shutter 2 is closed as shown in FIG. 5, the second space S2 is maintained in an atmosphere with a relatively low humidity.

  When the culture container 3 is transferred between the first space S1 and the second space S2, when the shutter 2 is opened, the air in the first space S1 passes through the open communication port 33. An attempt is made to flow into the upper space S21 of the second space S2. However, according to the automatic culture apparatus 1 according to the present embodiment, the communication port 33 is provided with the intake port 34, and as shown in FIG. The ambient atmosphere is aspirated from 34. Therefore, even when the shutter 2 is opened and the communication port 33 is opened, the hot and humid atmosphere in the first space S1 flows into the second space S2 and the temperature and humidity in the second space S2. It is prevented in advance.

  Moreover, according to the automatic culture apparatus 1 which concerns on this embodiment, the mechanism part for taking out the culture container 3 does not exist in the culture apparatus 4. FIG. That is, only a tray support member 4c for supporting the tray 7 in a state where the tray 7 is placed is provided in the culture apparatus 4, and all the mechanisms for taking out the culture container 3 are arranged outside the culture apparatus 4. The transport robot 5 is integrated. The transport robot 5 can be completely retracted to the outside of the door 4a of the culture apparatus 4 after the culture container 3 is taken in and out.

Therefore, when the door 4a is closed, there is no mechanism in the culture apparatus 4, and no dust is generated as generated by the operation of the mechanism. In addition, the inside of the culture apparatus 4 is maintained at a temperature of 37 ± 0.5 ° C., a humidity of 100%, a CO ₂ concentration of 5%, etc. The problem does not occur. Even when the door 4a is opened, only the tip of the hand 5c of the transfer robot 5 is inserted into the culture apparatus 4, and the rotation mechanism and the sliding mechanism substantially enter the culture apparatus 4. There is no. Therefore, intrusion of dust into the culture device 4 is suppressed, and the cleanliness inside the culture device 4 can be increased.
Note that the CO ₂ incubator culture apparatus 4, a multi-gas incubator, the incubator, such as the cool box, etc., may be constituted by one or a combination is used in the culture.

Furthermore, since the automatic culture apparatus 1 according to the present embodiment includes the air cleaning unit 23 above the central space S12 where the transfer robot 5 is installed, the automatic culture apparatus 1 is always in the central space S12 where the transfer robot 5 exists. Cleanliness is maintained. Therefore, even when the door 4a of the culture apparatus 1 is opened, it is possible to minimize the inflow of dust into the culture apparatus 1.
Therefore, according to the automatic culture apparatus 1 according to the present embodiment, it is possible to reduce the possibility that cells in culture are contaminated with dust and the like, and it is possible to culture healthy cells.

  In addition, this invention is not limited to the structure shown in the said embodiment. That is, the shape and number of the culture apparatus 1, the form and number of the transfer robot 5, the handling robot 10 and the dispensing robot 13, the form and number of various apparatuses, etc. are not limited at all and can be arbitrarily selected according to the application conditions. Can be set.

In addition to cytokines, for example, substances that contribute to growth such as concentrated platelets, BMP, FGF, TGF-β, IGF, PDGF, VEGF, HGF, and combinations of these should be employed as growth factors. It may be. In addition to penicillin antibiotics, any antibiotics such as cephem, macrolide, tetracycline, fosfomycin, aminoglycoside, and new quinolone can be used as the antibiotic.
Note that the automatic culture apparatus 1 according to the present embodiment is not limited to culturing bone marrow mesenchymal stem cells. You may culture the cell extract | collected from the various structure | tissue of the biological body, and the established cell line.

  Alternatively, the cells may be attached to a biological tissue filling material and cultured. As a biological tissue filling material, any material may be used as long as it is compatible with biological tissue instead of calcium phosphate, and a biologically absorbable material is more preferable. In particular, porous ceramics having biocompatibility, collagen, polylactic acid, polyglycolic acid, hyaluronic acid, or a combination thereof may be used. Further, a metal such as titanium may be used. The biological tissue filling material may be granular or block-shaped.

  Moreover, in the said embodiment, although the inlet port 34 was arrange | positioned in both the 1st space S1 side and the 2nd space S2 side of the communicating port 33, it may replace with this and may arrange | position in any one. . In addition, the suction pump is connected to the intake port 34. Instead, an intake blower is disposed in the lower space S22 of the second space S2, and when the shutter 2 is opened, After introducing the air into the lower space S22, for example, the air may be discharged to the outside of the housing 30 through a HEPA filter. Further, the position of the air inlet 34 is not limited to the lower part of the communication port 33 and may be arranged at an arbitrary position.

Furthermore, it is effective to start suction from the air inlet 34 before the shutter 2 is opened. By doing in this way, inflow of the hot and humid air from 1st space S1 to 2nd space S2 via the communicating port 33 is prevented more reliably.
In addition, the suction pump is operated in accordance with the opening of the shutter 2, but instead, as shown in FIG. 7, a door 35 for opening and closing the intake port 34 is provided to open and close the shutter 2. In addition, the door 35 may be opened and closed. The code | symbol 36 in the figure is a duct which connects the inlet port 34 and lower space S22 of 2nd space S2. By doing in this way, the air which is going to flow into the upper space S21 from the first space S1 through the communication port 33 using the intake air blower that forms the downdraft in the second space S2 It is possible to discharge from 34 to the lower space S22 through the duct 36 and to the external space via the HEPA filter.

It is a perspective view which shows the automatic culture apparatus which concerns on one Embodiment of this invention. It is a cross-sectional view which shows the automatic culture apparatus of FIG. It is a top view which shows the mode in the 1st space of the automatic culture apparatus of FIG. It is a perspective view which shows an example of the culture container used in the automatic culture apparatus of FIG. It is a perspective view which shows the communicating port which connects between the 1st space and 2nd space of the automatic culture apparatus of FIG. 1, and the shutter which interrupts | blocks this. It is a perspective view which shows a mode that the shutter of FIG. 5 is open | released and air is attracted | sucked by the inlet port. FIG. 6 is a partial longitudinal sectional view showing a modification of the intake port of FIG. 5.

Explanation of symbols

1 Automatic culture equipment 2 Shutter (open / close door)
3 Cultivation container 4 Cultivation equipment (culture room)
9 Conveyor (transport mechanism)
32 Processing Unit 33 Communication Port 34 Air Intake Port (Intake Unit)
S1 first space S2 second space

Claims (3)

A culture vessel containing cells is detachably accommodated, a first space for accommodating a culture chamber for culturing cells while maintaining predetermined culture conditions, and a predetermined treatment for the cells accommodated in the culture vessel A second space in which a processing section to be applied is disposed, and a transport mechanism for transporting the culture vessel via a communication port provided between the first space and the second space,
An automatic culture apparatus provided with an opening / closing door for opening and closing the communication port at the communication port, and an intake means for sucking air around the communication port when the door is opened.   The automatic culture apparatus according to claim 1, further comprising a control device that activates the intake means before the opening / closing door is opened.   The automatic culture apparatus according to claim 1 or 2, wherein the intake means includes an intake port on at least one of the first space side and the second space side of the communication port.

Images