WO2005061693A1 - Cell culture appartus - Google Patents

Abstract

It is intended to improve subculture performance and prevent contamination and cross-contamination by providing a cell culture apparatus which comprises: a culture means (15) of culturing cells which contains at least one cultivator (1) allowing control of its inner environment and having a medium inoculated with cells; a medium replacing means (9) of replacing the medium in the above cultivator; a cell measuring means (5) of measuring the conditions of the cells in the above cultivator; a cell inoculation means (11) of inoculating the cells in the above cultivator having been cultured by the above culture means into another cultivator having no cell inoculated; an air-tight chassis (17) for containing the above culture means, the above medium replacing means, the above cell measuring means and the above cell inoculation means which is provided with at least one opening/closing unit (19a) and (19b); and a cultivator carrying means (6) of carrying the above cultivator between the opening/closing unit of the above chassis and at least one of the above culture means, the above medium replacing means, the above cell measuring means and the above cell inoculation means.

Description

Cell culture equipment Technical field

 The present invention relates to a cell culture device, and more particularly, to a cell culture device for performing dairy culture. Background art

 When subculturing cells, change the culture medium in the incubator or use a new thread for subculture.

Complicated work such as re-seeding of cells into a medium is performed manually. In addition, the work associated with these subcultures must be performed by a skilled worker in order to suppress the occurrence of contamination and the like.

 As described above, the work involved in subculturing is currently performed manually, although it is complicated, and requires a skilled worker. Are difficult to work and difficult to implement.

 In particular, in the case of industrial cell culture that requires subculture, such as production of useful components using cultured cells and culture of stem cells for constructing tissue for regenerative medicine, the workability associated with subculture is important. Due to the poor quality, mass production is difficult, and improving the workability of subculture has been an issue.

 Japanese Patent Application Laid-Open No. 2001-275659 discloses an apparatus for automatically culturing cells in a closed environment by judging the timing of medium exchange and subculture by a camera in a culture tube to prevent contamination of cell culture. Is disclosed. Although this device employs a closed system that can reduce the risk of contamination by foreign substances such as microorganisms from the outside, the cultured cells are moved by a pump, which may cause damage to the cells.扱 う The risk of cross-contamination due to handling the cell suspension in the same closed system remained.

Japanese Patent Application Laid-Open No. 2002-262856 also discloses that the work load and contamination are reduced by removing the square culture tray from the incubator, replacing the culture medium, and returning the work to the incubator again, instead of manually. Nation and human error are reduced To disclose. However, when it was taken out of the incubator, it was opened to the open air, so that complete contamination was not achieved.

 An object of the present invention is to provide a culture apparatus capable of almost completely removing damage to cells, contamination and cross-contamination while maintaining the workability associated with the subculture. Disclosure of the invention

 The cell culture device of the present invention is a culture device that adjusts the internal environment and accommodates at least one incubator containing the medium in which the cells are seeded, and cultures the cells, and exchanges the culture medium in the incubator. Medium-exchange means for performing cell culture, cell state measuring means for measuring the state of cells in the incubator, and seeding the cells in the incubator cultured by the culture means into another incubator in which the cells have not been inoculated. A cell inoculating means, a transporting means for transporting the incubator, and an airtight housing containing the incubating means, the culture medium exchanging means, the cell state measuring means, the packet inoculating means, and the transporting means. The object is attained by providing the casing having at least one opening / closing section, and the transport means configured to transport the incubator at least between the opening / closing section and the cell seeding means.

 With this configuration, cells can be cultured, the medium in the incubator can be exchanged, the state of the cells in the incubator can be measured, and the state of the cells in the incubator can be renewed. Work associated with subculturing, such as re-seeding on the medium, can be performed with one device. In addition, it is necessary to suppress the generation of contamination during the work associated with these subcultures.However, all the work involved in these subcultures must be performed in a hermetically sealed housing in a state isolated from the external environment. And the generation of contamination can be suppressed. Therefore, it is not necessary to perform the complicated work associated with the subculture manually by a skilled worker, and the workability of the subculture can be improved. In addition, the culturing means has a configuration in which incubators are accommodated one by one and has a plurality of slots separated by partition walls. Contamination and cross-contamination due to the infection of the virus can be prevented.

Also, since no external force for feeding is applied to the cultured cells, the cells are damaged. I can't.

 Further, the housing has an inner partition that separates a space that houses the incubator that constitutes the culture means in the housing 1 ”and a space other than the space that houses the incubator. According to this structure, the space for accommodating the plurality of incubators constituting the culture means in the housing is provided as another space in the housing. In other words, it is preferable because the space for accommodating the incubator can be easily adjusted since the space for accommodating the incubator can be easily separated from the space where the medium is exchanged or seeded.

 In addition, the cell seeding means is configured to seed the cells in the incubator, which has measured the state of the cells, in another incubator if the cells have not been seeded, according to the result of the fh measurement by the cell state measuring means. Then, since the device automatically performs the subculture operation according to the measurement result of the cell state measuring means, the workability of the operation associated with the subculture can be further improved.

 In addition, if the cell state measuring means has a camera for photographing the incubator and is configured to measure the state of the cells by analyzing an image photographed by the camera, the operator can observe the image with a microscope or the like. It is preferable because the necessity of measuring the state of the cells can be eliminated, and the workability of the work associated with the subculture can be further improved.

 Further, the apparatus is provided with an identification information reading means for reading identification information attached to the incubator either before or after being carried into the housing. With such a configuration, information on the type and origin of the cells contained in the incubator for primary culture carried into the device, the state of culture in the incubator in the device, and the position of the incubator V, because it makes it easier to manage information about the incubator and information about the incubator.

 In addition, to the tower incubator to which the cells have been seeded by the cell seeding means, the relevant identification information attached to the identification information attached to the incubator from which the cells were collected, and the created identification information is added. Means. Such a configuration is preferable because it is possible to manage reports on an incubator in which cells have been replated for subculture.

In addition, it includes an ih command switch to command the suspension of culture in a specific incubator, and transports the incubator whose culture is suspended in response to the suspension instruction to the open / close section of the housing by transport means and discharges it from this open / close section. At the same time, from the incubator other than the incubator that stops the culture in response to the stop command in the enclosure, the identification information of the incubator that stops the culture in response to the stop command or related information The system has a culture suspending means for extracting identification information related to the identification information or an incubator to which the related identification information is attached, transporting the culture vessel to the opening / closing section of the housing by the transporting means, and discharging the culture vessel from the opening / closing section. With such a configuration, when an abnormality is found in the incubator, by instructing the culture to be stopped by the stop instruction switch, the original cells of the cells seeded in the incubator instructed to stop the culture are cultured. This is preferable because an incubator that has been inoculated, or an incubator in which cells have been replated and passaged from an incubator instructed to stop culturing can be excluded from the apparatus.

 In addition, in response to the determination of the abnormality by the cell state measuring means, the incubator in which the abnormality has been determined is transported to the opening / closing part of the housing by the transporting means and discharged from the opening / closing part, and the abnormality in the housing is determined. From the incubators other than the determined incubator, the incubator with the identification information related to the incubator in which the abnormality was determined or the related identification information or with the related identification information is extracted, and the housing is mounted on the housing by the transportation means. It is configured to have a culture stopping means for transporting to and opening from the opening / closing section. With such a configuration, when an abnormality is detected in the incubator, the incubator in which the abnormality is detected and the incubator related to the incubator can be automatically removed from the apparatus, and the work involved in subculturing can be performed. This is preferable because the workability of the method can be further improved. Furthermore, when there is an incubator containing a medium in which cells are seeded in the housing, at least one of an alarm and holding the open / close portion of the housing in a closed state allows the incubator to be mounted in another incubator. The incubator has a means for preventing carry-in. With such a configuration, in order to prevent cross contamination, when one cell culture device is used for culturing one cell, an incubator containing another cell is erroneously carried into the device. This is preferable because it is possible to prevent that.

In addition, based on the identification information recognized by inputting or reading the identification information attached to the incubator before being carried into the housing, the cells in the incubator with this identification information can be used in the incubator inside the housing. When it is detected that the cells have different origins, the alarm and / or the opening / closing part of the housing is kept closed to prevent the incubator containing cells of different origins from being brought into the housing. The incubator is provided with an incubator blocking means. With such a configuration, if one cell culture device is used for culturing cells of one origin to prevent cross contamination, the incubator containing cells of different origins may be incorrectly placed in the device. It is preferable because it can be prevented from being carried into the inside. Further, if the air conditioner is provided with a pressure adjusting means for adjusting the air pressure inside the housing to a more positive pressure than the air pressure around the housing, the opening / closing portion of the housing can be, for example, an air-lock type, so that an atmosphere outside the housing can be used. It is not necessary to make the structure hard to flow into the body, and the structure of the opening / closing part of the housing can be simplified, which is preferable. Furthermore, if the pressure in a space other than the space accommodating the incubator is lower than the space accommodating the incubator constituting the culture means in the housing, contamination to the incubator can be easily prevented.

 Further, the apparatus has an ultraviolet irradiation means for irradiating at least a space in the housing where cells are seeded by the cell seeding means and a space where the medium is exchanged by the medium exchange means. When at least one of the space where the cells are seeded by the seeding means and the space where the medium is exchanged by the medium exchange means is provided with an incubator, irradiation of ultraviolet rays is stopped. Such a configuration is preferable because microorganisms in the space where the culture medium in the incubator is exposed to the air in the housing can be killed without affecting the culture. Brief Description of Drawings

FIG. 1 is a block diagram showing a schematic configuration and operation of an embodiment of a cell culture device to which the present invention is applied. FIG. 2 is a cross-sectional view showing a specific configuration of one embodiment of the Itoda spore culturing apparatus to which the present invention is applied. FIG. 3 is a block diagram showing a schematic configuration of a connection state between a controller and each operating portion of one embodiment of the cell culture device to which the present invention is applied. FIGS. 4A and 4B are perspective views showing the appearance of an embodiment of a cell culture device to which the present invention is applied, wherein FIG. 4A shows a state in which the cell culture device is installed alone, and FIG. It is a figure showing the state where several cell culture devices were connected and installed. FIG. 5 is a flowchart showing a schematic operation of an embodiment of the cell culture device to which the present invention is applied. FIG. 6 is a block diagram showing a schematic configuration and operation of a modification of the cell culture device to which the present invention is applied. FIG. 7 is a block diagram showing a schematic configuration and operation of another modification of the cell culture device to which the present invention is applied. FIG. 8 is a block diagram showing a schematic configuration and operation of still another modified example of the cell culture device to which the present invention is applied. BEST MODE FOR CARRYING OUT THE INVENTION An embodiment of the present invention will be described. Hereinafter, an embodiment of a cell culture apparatus to which the present invention is applied will be described with reference to FIGS. FIG. 1 is a block diagram showing a schematic configuration and operation of a cell culture device to which the present invention is applied. FIG. 2 is a cross-sectional view showing a more specific configuration of the cell culture device of the present embodiment. FIG. 3 is a block diagram showing a schematic configuration of a connection state between a controller of the cell culture device of the present embodiment and each operating portion. FIG. 4 is a perspective view showing ^ m of the cell culture device to which the present invention is applied,

(a) is a diagram showing a state where a single cell culture device is installed, and (b) is a diagram showing a state where a plurality of cell culture devices are connected and installed. FIG. 5 is a flowchart showing a schematic operation of a cell culture apparatus to which the present invention is applied.

 As shown in FIG. 1, the cell culture device according to the present embodiment includes an ID reader 3 for reading identification information, ie, ID, attached to the incubator 1 attached to the incubator 1, and measures the state of the cells. Dividing means that forms part of the cell measuring means 5 and the transporting means 6 and selects the transport destination of the incubator 1 by branching the transporting path of the incubator 1, a medium for exchanging the culture medium in the incubator 1 Exchange means 9, Cell seeding means for disseminating and distributing cells in incubator 1 into medium in another incubator 1, Related identification information related to the identification information read by ID reading means 3, that is, related ID It is provided with a related ID assigning means 13 for assigning to the incubator 1 in which cells are seeded by the cell seeding means 11 and a culturing means 15 which is an incubator.

 Furthermore, the cell culture device of the present embodiment includes the ID reading means 3, the cell measurement means 5, the transport means 6, the flow dividing means 7, the medium exchange means 9, the cell seeding means 11, the related ID assigning means 13, and the culture. The means 15 and the like are housed in an airtight housing 17. In the cell culture device of the present embodiment, a controller 21 for controlling the operation of the cell culture device is provided outside the housing 17.

The housing 17 has a cell carry-in port 19a and a cell carry-out port 19b serving as an opening / closing unit. The transport means 6 transports the incubator from the cell entrance 19a of the housing 17 to the flow dividing means 7 through the ID reading means 3 and the cell measuring means 5 in order. Further, in the transport means 6, the transport path of the incubator is branched by the splitting means 7, and in one route, the splitting means 7 passes through the medium exchange means 9 or the cell seeding means 11 and the related ID assigning means 13 in order. Then, the incubator 1 is transferred to the culture means 15. In another route, the culture that has been The incubator 1 is transported to the cell outlet 19b, and the incubator 1 to be discarded when an abnormality or the like has occurred is transported to the cell outlet 19b. On the other hand, the transfer means 6 transfers the incubator 1 from the culture means 15 to the cell measurement means 5.

 The culture means 15 is separated from other spaces in the housing 17 by an internal partition 23. The inner bulkhead 23 is provided with an inner carry-in port 25a and an inner carry-out port 25b serving as inner opening / closing portions. Therefore, the transport means 6 transports the incubator 1 from the medium exchange means 9 to the inner loading port 25a, or from the related ID assigning means 13 to the inner loading port 25a, and also transfers the incubator 1 from the inner loading port 25b to the cell measuring means 5. Convey incubator 1. Further, the culturing means 15 includes a carbon dioxide sensor 27 for detecting the concentration of carbon dioxide inside the culturing means 15, and a carbon dioxide cylinder 29 connected at one end, and a culturing means for the culturing means 15 at the other end. It has a carbon dioxide supply line 31 communicating with the culture space accommodating the vessel 1. An electromagnetic valve 33 is provided in the carbon dioxide supply line 31, and the electromagnetic pulp 33 and the carbon dioxide concentration sensor 27 are electrically connected to the controller 21 through a wiring 35. I have. The culturing means 15 is provided in a space constituting the culturing means 15 detected by the controller 21 with the carbon dioxide concentration sensor 27, that is, a carbon dioxide supply pipe according to the carbon dioxide concentration in the culturing space accommodating the incubator 1 of the culturing means 15. By controlling the flow rate by the electromagnetic valve 33 of the passage 31, the carbon dioxide concentration in the culture space accommodating the incubator 1 of the culture means 15 can be maintained within a certain range. Further, the culturing means 15 includes a heater such as a heater, a temperature sensor, and the like (not shown), and can maintain the temperature in the culturing space accommodating the incubator 1 of the culturing means 15 within a certain range. it can. As described above, the culturing unit 15 is a carbon dioxide gas incubator, and can adjust the carbon dioxide concentration and temperature in the culturing space accommodating the incubator 1 of the culturing unit 15, that is, the internal environment. Although not shown in FIG. 1, the culturing means 15 is a slot for accommodating the incubator 1 in the culturing space of the culturing means 15, and the culturing means is connected between the slot and the inner carry-in port 25a or the inner carry-out port 25. It has a transportation means in the culture means for transporting 1 and the like.

On the other hand, in a space other than the culture space of the culture means 15 in the housing 17, a carbon dioxide supply pipe 31 provided with an electromagnetic valve 33 is connected similarly to the culture means 15. A carbon dioxide concentration sensor 27 for detecting the carbon dioxide concentration in the space other than the culture space of the culture means 15 in {7} is provided. For this reason, the culture The concentration of carbon dioxide in a space other than the culture space of the step 15 can be adjusted to the same state as in the culture space of the culture means 15, and when the incubator 1 is loaded or unloaded into the culture space of the culture means 15. Furthermore, even if the inner carry-in port 25a and the inner carry-out port 25b provided in the inner partition 23 that defines the culture means 15 are opened, the internal environment of the culture space of the culture means 15, that is, the culture environment can be hardly changed.

 The controller 21 includes an ID reading unit 3, a cell measuring unit 5, a transporting unit 6, a flow dividing unit 7, a medium exchange unit 9, a cell seeding unit 11, and a related ID providing unit 13, which are surrounded by a broken line in FIG. It is electrically connected to the transport means 6 and the like via the wiring 35, and the ID reading means 3, the cell measuring means 5, the transport means 6, the flow dividing means 7, the medium exchange means 9, the cell seeding means 11, the related IDs The control unit that controls the operation of the application unit 13 and the culture unit 15 is a part of each unit. In addition, the controller 21 is a monitor for displaying various information such as input devices such as a keyboard and a mouse, information about control and information about a culture state such as a result of measurement by the cell state measuring means 5, and a process of culture. And storage means for storing various parameters, identification information, related identification information, and the like in each step of the culture.

 A more specific configuration of such a cell culture device of the present embodiment will be described below. In the cell culture apparatus of the present embodiment, as shown in FIG. 2, one opening / closing section 19 serving as the cell entrance 19 a and the cell exit 19 b shown in FIG. 1 is provided on one surface of the side wall of the housing 17. The space on the side wall provided with the opening / closing section 19 in the housing 17 is a medium exchange and cell seeding section 37, and an ID reader 3, cell measurement means 5, and transport means are provided in the space for the medium exchange and cell seeding section 37. 6, each device constituting the flow dividing means 7, the medium exchange means 9, the cell seeding means 11, the related ID assigning means 13, and the like. On the other hand, the space on the side wall opposite to the side provided with the opening / closing portion 19 in the housing 7 is referred to as a culture portion 39, and the space serving as the culture portion 39 accommodates the incubator 1 and accommodates the incubator 1. This is a culture space for culturing cells seeded in the medium. The internal partition 23 separates the medium exchange and cell seeding section 37 from the culture section 39. The inner partition wall 23 is provided with one inner opening / closing section 25 which also serves as the inner carrying-in port 25a and the inner carrying-out port 25b shown in FIG.

The opening / closing portion 19 provided in the housing 17 closes the inside of the housing 17 when closed, and a shutter 43 covering the opening 41 of the opening / closing portion 19 of the housing 17, and an upper edge portion of the shutter 43. It is composed of a fixed wire 45, a shirt motor 47 fixed above the opening 41 on the wall of the housing 17, and a pulley 49 fixed to the rotating shaft of the shutter motor 47 and winding the wire 45. I have. The shutter motor 47 of the opening / closing section 19 is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG. As shown in FIG. 2, the inner opening / closing portion 25 provided in the inner partition wall 23 includes a shirt 53 covering an opening 51 formed in the internal partition 23, a wire 55 fixed to an upper edge portion of the shirt 53, The shutter 23 includes a shutter motor 57 fixed to the upper part of the opening 51 of the partition wall 23, a pulley 59 fixed to the rotation shaft of the shutter motor 57 and winding the roller 55.

The shutter 43 and the shutter 53 of the opening / closing section 19 and the inside opening / closing section 25 are opened and closed as indicated by arrows A and C in FIG. 2 by driving the shutter motor 47 and the shutter motor ₅₇ , respectively. The shutter motor 57 of the opening / closing section 25 is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG.

 Between the opening / closing section 19 and the inside opening / closing section 25 in the housing 17, a first belt conveyor 61 serving as a conveying means 6 for conveying the incubator 1 from the opening / closing section 19 to the inside opening / closing section 25 is provided. I have. The first belt conveyor 61 is hung between rollers 61a installed on the opening / closing section 19 side, rollers 61b installed on the inner opening / closing section 25 side, and rollers 61a and rollers 61b arranged horizontally apart. Belt 61c, and a roller motor 61d connected to the roller 61a to rotate the roller 61a. The first belt conveyor 61 conveys the incubator 1 in the horizontal direction by driving the roller motor 61d, as shown by the arrow C in FIG. As shown in FIG. 3, the roller motor 61d of the first belt conveyor 61 is connected to the 1/0 port 21a of the controller 21 via the wiring 35.

Above the opening / closing portion 19 side end of the first belt conveyor 61, an incubator 1 serving as an ID reader constituting ID reading means 3 for reading identification information attached to the incubator 1 placed on the first belt conveyor 61 is provided. A par code reader 63 is provided for reading a bar code corresponding to the identification information pasted on the bar code. That is, the identification information is obtained by attaching a label printed with a barcode corresponding to the identification information to the incubator 1. It is attached to. The LED 63a and the decoder 63b of the barcode reader 63 are connected to the controller 21 via the wiring 35 as shown in FIG.

 Above the end of the opening / closing section 19 side of the first belt conveyor 61, on the inside opening / closing section 25 side of the barcode reader 63, the cell state measuring means 5, the medium exchange means 9, and the cytotoxic seed means 11 are configured. A turntable 65 is provided. The turntable 65 includes a disk-shaped table 65a, a turntable motor 65b installed on the lower surface side of the table 65a, and horizontally rotating around the center of the table 65aO. On the upper surface of the table 65a, the main part la containing the culture medium of the incubator 1, that is; : 咅 A not-shown depression 1S large enough to receive the main part la after removing the lid part lb of the culture vessel 1S is formed at predetermined intervals in the circumferential direction. Such a turntable 65 is rotated by the drive of the turntable motor 65b as indicated by the arrow K in FIG. Then, the turntable motor 65b of the turntable 65 is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG. Above the turntable 65, corresponding to the position of the turntable 65, a piveting mechanism and an imaging unit 67 that constitute the cell state measuring means 5, the medium exchange means 9, and the cell seeding means 11 together with the turntable 65. Is provided. The piving mechanism and the photographing unit 67 are fixed to a portion corresponding to the position of the turntable 65 on the inner surface of the ceiling of the housing 17 and are supported by the support member 67a that is suspended toward the turntable 65. It is fixed to the inner surface. The support member 67a is provided with a plurality of pinions 67b arranged in the vertical direction. A rack 67c extending upward is connected to the pinion 67b by engaging with the pinion 67b.

 A pipette base rotation motor 67d is fixed to the lower end of the rack 67c. The rotation axis of the pipe base rotation motor 67d is fixed to the center of a disk-shaped pipette base 67e located at the bottom IJ of the pipe base rotation motor 67d. One of the pinion pins 67b is connected to the rotating shaft of a pipette base elevating motor 67f. The pipe base rotary motor 67d and the pipe base elevating motor 67f are arranged as shown in FIG. Next, the controller 21 is connected to the I / O port 21a of the controller 21 via the wiring 35, and is connected.

At the center of the lower surface of the pipe base 67e, as shown in FIG. The microscope CCD camera unit 67g that constitutes 5 is attached. At a position facing the peripheral portion of the pipe base 67e, a medium exchange pipe nozzle 67h constituting the medium exchange means 9 and a cell seeding pipe nozzle 67i constituting the cell seeding means 11 are attached. I have. The microscope CCD camera unit 67g is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG. Such a pivet mechanism and the photographing section 67 are driven by a pit base rotation motor 67d to rotate a pit base 67e as indicated by an arrow L in FIG. 2, and driven by a pit base lifting motor 67f. The pivot 67e moves up and down as indicated by the arrow M in FIG. As shown in FIG. 2, the turntable 65, the pitting mechanism, and the imaging unit 67 are composed of a microscope CCD camera unit 67g, a medium replacement pit nozzle 67h, and a cell seeding pit nozzle. One of the nozzles 67i and one of the nozzles is installed in a positional relationship so as to come on the incubator 1 mounted on the table 65a of the turntable 65. In addition, the pipe nozzle 67h for medium exchange and the pipe nozzle 67i for cell seeding were arranged such that the required pipe nozzle was placed on the table 65a of the turntable 65 by rotating the pipe base 67e. Come on incubator 1. Further, the pipe base 67e may be provided with a nozzle for adding a chemical such as trypsin for detaching cells from the incubator, in addition to the medium replacement pipe nozzle 67h and the cell seeding pipe nozzle 67i. it can.

One end of a liquid sending tube 69 is connected to the medium replacement pipe nozzle 67h. The other end of the liquid sending tube 69 is connected from the inside of the housing 7 to an aspirator unit 71 provided on the side wall portion above the opening / closing unit 19 outside the housing 17. The aspirator section 71 has a downwardly directed discharge nozzle 71a, and a tray 73 for receiving the discharged medium is provided below the discharge nozzle 71a outside the housing 17. A pump 77 connected to an aspirator section 71 via a tube 75 is provided on the outer ceiling surface of the housing 17. The pump 77 is capable of switching between suction and delivery. At the time of suction, the aspirator 71 is set to a negative pressure, and the medium in the incubator 1 is suctioned by the medium replacement pipette nozzle 67h and discharged to the tray 73. Further, the aspirator section 71 internally has a flow passage (not shown) for switching a flow path electrically connected to the controller 21. The pulp is used to switch the flow path, and this time, a new sterilized medium is sucked from a medium tank (not shown) installed outside the housing 17. And inject it into the incubator 1 from the medium exchange pipe nozzle 67h.

 One end of a liquid sending tube 79 is connected to the cell seeding pipe nozzle 67i. The other end of the liquid feeding tube 79 is connected to the pump 81 provided on the ceiling surface outside the housing 17 from the inside of the housing 17. The pump 81 is capable of switching between suction and delivery. During the suction, the fine JI packet in the incubator 1 is sucked into the cell seeding pipe nozzle 67i. Remove the aspirated cells. In this way, the cells in the incubator 1 that have reached confluence can be seeded in a new culture medium in the incubator 1 and the cells can be passaged. The pumps 77 and 81 are connected to the IZO port 21a of the controller 21 via the wiring 35 as shown in FIG. In order to irradiate the space where the pitting mechanism and the imaging unit 67 and the turntable 65, etc. constituting the cell state measuring means 5, the medium exchange means 9, and the cell seeding means 11 are installed, a housing is provided. An ultraviolet lamp 82 is provided on the ceiling surface in the vicinity of the pivetting mechanism 17 and the photographing section 67 in 17. The ultraviolet lamp 82 is connected to the IZO port 21a of the controller 21 via the wiring 35 as shown in FIG. When the incubator 1 in which cells were seeded in the medium exchange and cell seeding section 37 is present, the controller 21 turns off the ultraviolet lamp 82, and the cells are seeded in the medium exchange and cell seeding section 37. If there is no incubator 1, the ultraviolet lamp 82 is turned on.

Above the first belt conveyor 61 between the turntable 65 and the inner opening / closing section 25, as shown in FIG. 2, a second belt conveyor 83 is provided. The second conveyor / reto conveyor 83 includes a roller 83a installed on the turntable 65 side, a roller 83b installed on the inner opening / closing section 25 side, and a roller 83a and a roller 83 arranged horizontally apart. It is formed by a belt 83c that is hung, a roller motor 83d that is connected to the roller 83a, and drives the roller 83a to rotate. The second belt conveyor 83 conveys the incubator 1 in the horizontal direction as indicated by the arrow F in FIG. 2 by driving the roller motor 83d. Then, as shown in FIG. 3, the roller motor 83d of the second veneto conveyor 83 connects the wiring 35 to the IZO port 21a of the controller 21. Connected through.

 Above the turntable 65 side of the second belt conveyor 83, as shown in FIG. 2, means for removing the lid part lb of the incubator 1 on the second belt conveyor 83, and further, An incubator gripping arm 85 is provided as a part of the transport means 6 for transporting the main body la of the incubator 1 with the lid lb coming to the end portion on the turntable 65 side removed to the turntable 65. The incubator gripping arm 85 is located at the position corresponding to the turntable 65 side of the second belt conveyor 83 and the portion of the turntable 65 on the second belt conveyor 83 on the inner surface of the ceiling of the housing 17. A plurality of pinions 85a installed in a direction along the direction in which the conveyor 83 extends, a horizontal rack 85b extending in the horizontal direction along the direction in which the second belt conveyor 83 extends along the plurality of pinions 85a, and It is fixed to the inner surface of the ceiling of the housing 17 by a hanging support member 85c fixed to the horizontal rack 85b.

 The support member 85c is provided with a plurality of pinions 85d arranged in the vertical direction. When the pinions 85d squirt, the vertical racks 85e extending in the vertical direction are connected to each other. One of the plurality of pinions 85a and one of the plurality of pinions 85d is connected to a rotating shaft of an arm slide motor 85f and an arm elevating motor 85g, respectively. A gripping motor 85h is fixed to a lower end of the vertical rack 85e. The gripping motor 85h is fixed to a gripping portion 85i that opens and closes as indicated by the solid and broken lines in FIG. 2 by the rotation of the gripping motor 85h. In the incubator gripping arm 85, the gripping portion 85i is opened and closed as shown by the arrow G in FIG. 2 by driving the gripping motor 85h, and the arrow H in FIG. 2 is driven by the arm lifting motor 85g. As shown by, the grip portion 85i moves up and down, and the grip portion 85i moves in the horizontal direction by the drive of the arm slide motor 85f as shown by the arrow J in FIG. The arm slide motor 85f, the arm lifting / lowering motor 85g, and the gripping motor 85h are connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG.

Above the second belt conveyor 83, near the inner opening / closing section 25 within the movable range of the incubator gripping arm 85 in the horizontal direction, as shown in FIG. A lid placement shelf 87 for placing the lid portion lb removed by 85 is provided. Furthermore, above the inner opening / closing section 25 side end of the second belt conveyor 83 Constitutes related ID assigning means 13 for assigning related identification information to the incubator 1 at the end of the inner opening / closing section 25 side of the second belt conveyor 83, and a bar code corresponding to the related identification information is labeled. A bar code printing and sticking device 89 is provided for printing and sticking the bar code printed label to the target incubator 1. Then, the barcode printing / pasting device 89 is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG.

 As shown in FIG. 2, the inside of the culturing section 39 constituting the culturing means 15 has a side wall facing the side where the opening / closing section 19 is provided in the housing 17, that is, a side wall side facing the inside opening / closing section 25. A plurality of slots 91 for accommodating the incubator 1 are provided. The plurality of slots 91 are formed in a plurality of shelves arranged in the vertical and horizontal directions. The plurality of slots arranged in the vertical and horizontal directions are separated by a partition 93 between adjacent slots 91. This prevents the incubator 1 housed in the combined slot 91 from contacting. In addition, since FIG. 2 is a cross-sectional view, a plurality of slots 91 arranged in the vertical direction are not shown, and the partition wall 93 is also provided with the bottom of the slots 91 for partitioning the plurality of slots 91 arranged in the vertical direction. The septum is not shown. However, actually, a plurality of slots 91 are arranged in the horizontal direction, and the adjacent slots 91 are separated by a partition wall 93 serving as a side wall of the slot 91. .

 The plurality of slots 91 have an in-slot belt conveyor 95 for moving the incubator 1 between the inner side opening / closing portion 25 side end of the slot 91 and the side wall side end facing the inner side opening / closing portion 25. The belt conveyor 95 in the slot is provided with rollers 95a installed on the inner opening / closing section 25 side, rollers 95b installed on the side wall side of the housing 17 facing the inner opening / closing section 25, and is horizontally spaced. It is formed by a belt 95c hung between the rollers 95a and 95b, and a roller motor 95d connected to the roller 95a to rotate the roller 95a. Such a slot belt conveyor 95 conveys the incubator 1 in the horizontal direction as indicated by the arrow D in FIG. 2 by driving the roller motor 95d. Then, the roller motor 95d of the slot belt conveyor 95 is connected to the IZO port 21a of the controller 21 via the wiring 35 as shown in FIG.

The gap between the opening at the inside opening / closing section 25 side end of the slot 91 and the inside opening / closing section 25 is as shown in FIG. As shown in FIG. 1, one third belt conveyor 97 is provided. The third belt conveyor 97 was hung between the rollers 97a installed on the inner opening / closing section 25 side, the rollers 97b installed on the slot 91 side, and the rollers 97a and 97b arranged at a horizontal interval. The belt 97c is formed by a roller motor 97d connected to the roller 97a and driving the roller 97a to rotate. Furthermore, the third belt conveyor 97 has a roller 97a, a roller 97b, a belt 97c, and a base 97e that supports a roller motor 97d and the like. Such a third belt conveyor 97 conveys the incubator 1 in the horizontal direction as indicated by the arrow D in FIG. 2 by driving the roller motor 97d. Further, the roller motor 97d of the third belt conveyor 97 is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG.

 In addition, as shown in FIG. 2, an elevating unit is provided in the culturing unit 39 constituting the culturing unit 15 so as to extend in the up and down direction near the internal partition wall 23 and move the third belt conveyor up and down in the vertical direction. A mechanism section 99 is provided. The elevating mechanism 99 is provided between the roller 99a provided on the lower side in the culture unit 39, the roller 99b provided on the upper side in the culture unit 39, and the rollers 99a and the rollers 99b arranged at intervals in the vertical direction. And a roller motor 99d that is connected to the roller 99a and drives the roller 99a to rotate. The base 97e of the third belt conveyor 97 is fixed to the belt 99c. Such a lifting mechanism 99 moves the third belt conveyor 97 in the vertical direction as indicated by an arrow E in FIG. 2 by driving the roller motor 99d. Then, the roller motor 99d of the lifting mechanism 99 is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG. Note that the third belt conveyor 97 and the elevating mechanism 99 constitute the flow dividing means 7 in FIG.

 Inside the medium exchange and cell seeding section 37 and the culture section 39 of the housing 17, diacid carbon nozzles 101 for supplying carbon dioxide into the respective spaces are provided. One end of a carbon dioxide supply pipe 31 is connected to the carbon dioxide nozzle 101. Then, the electromagnetic pulp 33 provided in the carbon dioxide supply pipe 31 is connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG.

Further, air nozzles 103 are provided in the medium exchange and cell seeding section 37 and the culture section 39 of the housing 17, respectively. The air nozzle 103 has an air supply line 105 One end is connected. The other end of the air supply pipe 105 is connected to an air supply source such as a pump that sucks air and sends air. Although not shown, the air supply line 105 or the air supply source is provided with a means for purifying air such as a HEPA filter or an ULPA filter, and the purified air is supplied to the culture medium. It is supplied into the exchange and cell seeding unit 37 and the culture unit 39. Further, a pressure sensor (not shown) is provided in each of the medium exchange and cell seeding unit 37 and the culture unit 39. The air supply line 105 is provided with an electromagnetic valve 107 for adjusting a flow rate of supplied air.

 Then, the electromagnetic valve 107 provided in the air supply conduit 105 and a pressure sensor (not shown) are connected to the I / O port 21a of the controller 21 via the wiring 35 as shown in FIG. As a result, the controller 21 adjusts the opening of the electromagnetic valve 107 according to the pressure detected by the pressure sensor, and supplies clean air into the medium exchange and cell seeding unit 37 and the culture unit 39 to supply each air. The pressure in the space is adjusted so that the inside of the medium exchange and cell seeding unit 37 and the culture unit 39 is at a more positive pressure than the outside of the housing 17.

 As shown in FIG. 3, the controller 21 is formed of a computer including a CPU 107 and a memory 109 serving as a storage unit, and includes an input device and a monitor as described above. The IZO port 21a of the controller 21 has a variable resistor 111 for setting the culturing time in addition to the above-mentioned devices, a case where an abnormality occurs in the cultured cells, and a housing for cells of different origins and types.警報 When the setting to prohibit loading into 7 is set, the alarm lamp 113 and the alarm speaker 115 are used to alert the operator when trying to bring cells of different origins or types into the housing 17. In addition, a culture stop command switch 1-7 for giving a command to stop the culture when an abnormality occurs in the cultured cells is connected via the wiring 35. The alarm lamp 113 and the alarm sound speaker 115 are connected to the I / O port 21a of the controller 21 via the amplifier 119 and the amplifier 121, respectively.

Here, the controller 21 has a function of retrieving a culture protocol such as a culture process and conditions set and stored in advance based on the identification information corresponding to the barcode read by the percode reader 63. Have. Also, slot identification information such as a unique number is assigned to each slot 91, and the controller 21 controls the bar code dolly. Identifier corresponding to the barcode read by the reader 63 or the related identification information corresponding to the barcode affixed by the barcode printing and pasting device 89, and the incubator 1 provided with the identification information or the related identification information. Is stored in association with the slot identification information attached to the slot 91 accommodating the. The controller 21 manages the culturing process and state of the incubator 1 in the housing 17 based on the slot identification information attached to the slot 91. Therefore, it is not necessary to transport the incubator 1 to the position of the bar code reader 63 each time identification information is required.

 Further, the controller 21 has a function of praying an image captured by the microscope CCD camera unit 67g, judging a confluent state from a cell area or the like, and calculating a cell number. Further, when the operator determines an abnormality based on the image of the incubator 1 displayed on the monitor and instructs the suspension of the culture using the stop command switch 117, the controller 21 starts the culture performed at that time. Conveyor 1 is transported to the opening / closing section 19 of the housing # 7, discharged from the opening / closing section 19 to the outside of the housing 17, and the related identification information related to the identification information of the incubator 1 or the related identification information taken at that time. The incubator 1 marked with is extracted from the slot 91 of the culturing section 39, and is transported to the opening / closing section 19 of the housing 17 and discharged from the opening / closing section 19, thereby serving as a culture stopping means. In addition, the culture suspension means consisting of 21 controllers determines the suspension of culture when abnormalities are detected from the results of analyzing images taken with a microscope microscope 67g CCD camera cutout. Similarly, the incubator 1 in which the abnormality was detected, the incubator 1 of the original cell in which the cells were subcultured in the incubator 1 in which the abnormality was detected, and the incubator 1 which was subcultured from the incubator 1 in which the abnormality was detected It can also be set to discharge the incubator 1 for the cells that have been removed.

Further, when the cell culture device needs to culture only one cell, the controller 21 can be set, if the incubator 1 containing the medium in which the cells are seeded in the housing 17 is set, for example. By locking the shutter motor 47 so that it cannot be driven, or by inserting a pin into the hole formed in the shutter motor 47, the shutter 43 of the opening / closing section 19 of the housing # 7 is kept closed. However, it functions as an incubator carrying-in prevention means for preventing carrying-in of another incubator 1. If the operator operates a switch or the like that opens the shutter 43 to load the incubator 1, the controller 21 turns on the alarm lamp 113 and sounds an alarm sound from the alarm speaker 115. It also has the function of notifying that the culture is being performed.

 If a barcode reader is provided outside the housing 17 or the identification information attached to the incubator 1 can be input from the input devices of the controller 21, if the cells are of the same origin, If the incubator 1 containing the medium in which the cells are seeded is present in the housing 17 depending on the setting, if the incubator 1 can be carried into the housing # 7, the identification information read or input and the housing Γ Compare the identification information of the incubator 1 that has already been loaded into the Γ7, and if the result of the comparison indicates that no cells of the same origin appear, hold the shutter 43 of the opening / closing section 19 of the housing 17 closed. It also functions as an incubator loading prevention means for preventing the loading of another incubator 1.

 In the present embodiment, the incubator 1 containing a medium not seeded with the cells to be used for subculture is configured to be housed in the slot 91 in the culture unit 39, but is transferred in the housing 17 It is also possible to adopt a configuration in which a portion for accommodating an incubator 1 containing a medium not seeded with cells to be used in the alternative is provided. Alternatively, the housing # 7 may be connected to an autoclave or the like, and the incubator 1 containing a medium not seeded with sterilized cells may be carried into the housing 17 via the autoclave. .

 In addition, in the cell culture apparatus of the present embodiment, as shown in FIG. 4A, one opening / closing section 19 serving as a cell carrying-in port 19a and a cell carrying-out port 19b is provided on one side wall 123 of the housing 17. The outer surface of the side wall 125 connected to the side wall 123 provided with the opening / closing portion 19 at an angle of 90 degrees is formed flat. For this reason, as shown in FIG. 4 (b), the cell culture device of the present embodiment can be set up by arranging a plurality of cell culture devices by bringing the outer surfaces of the side walls 125 into contact with each other. Therefore, since one cell culture device is used for subculture of one cell, the installation space of the cell culture device can be reduced even when it is necessary to install a plurality of cell culture devices.

The operation of the cell culture device having such a configuration and the features of the present invention will be described. Here, an example will be described in which a single cell culture device is used to passage cells from one incubator to increase the number of cells in order to prevent cross contamination. When the operator turns on a switch or the like for instructing the shutter 43 of the opening / closing section 19 to be opened with input devices or the like of the controller 21, as shown in FIG. To determine whether the cells have already been loaded into the cell, that is, whether the cells have already been cultured. SI). If it is determined in step SI that the cells have already been carried into the housing 17, the alarm function is turned on, the alarm lamp 113 is turned on, an alarm sound is emitted from the alarm speed 115, and the opening / closing section 19 is opened. Lock so that the shutter 43 does not open (step S3). On the other hand, if it is determined in step S1 that no cells have been introduced into the housing 17, the ultraviolet lamp 82 is turned off (step S5), and then the shutter 43 of the opening / closing section 19 is opened (step S7).

 Here, the operator carries in the incubator 1 seeded with cells to be propagated by performing primary culture and subculture, from the shutter 43 of the opening / closing section 19, and places it on the first belt conveyor 61. At this time, a label printed with a barcode corresponding to the identification information is attached to the incubator 1 to be carried in. Then, a switch or the like for instructing the start of the culture is turned on by the input devices of the controller 21 or the like. Thereby, the controller 21 closes the shutter 43 of the opening / closing section 19 (Step S9). Then, the first belt conveyor

The par code printed on the label attached to the incubator 1 placed on the 61 is read by the par code reader ₆₃ to obtain the identification information of the incubator 1 (step Sll). The controller 21 searches and obtains information about the cells to be cultured, which is set and stored in advance, based on the identification information of the incubator 1 obtained here, and a culture protocol such as culture procedures and conditions. Culture is performed according to the culture protocol obtained here.

 After step S11, the controller 21 transfers the incubator 1 to the turntable 65 using the first belt conveyor 61, the third belt conveyor 97, the second belt conveyor 83, and the incubator gripping arm 85, which are the transfer means 6. It is transported, and the incubator 1 is placed on the turntable 65. Then, the cell state is measured from the image of the incubator 1 photographed by the pipetting mechanism and the microscope CCD camera unit 67g of the photographing unit 67 (step S13, step S15). In step S13, the color, shape, size, presence or absence of contamination, and the like of the cell are measured. If it is determined in step S13 that there is no abnormality such as death of the cell or occurrence of contamination, the controller 21 determines in step S15 whether the cell has reached the confluent state by measuring the cell density and the like.

Here, since the culture has not been performed yet, the controller 21 proceeds to step S15. It is judged from the state of the cells that the confluent has not been reached. Further, the incubator 1 is conveyed and accommodated in the slot 91 of the culture section 39 by the third belt conveyor 97, and primary culture is performed under preset conditions (step S17). At this time, the controller 21 stores the identification information of the incubator 1 housed in the slot 91 in association with the slot identification information of the slot 91. In step S17, the incubator 1 is transported from the slot 9 to the turntable 65 for a preset time, and the pipetting mechanism constituting the medium exchange means and the medium exchange pipe nozzle 67h of the imaging unit 67 are used. Remove the old medium and replace with new medium.

 When the preset incubation time has elapsed (step S19), the controller 21 extracts the incubator 1 again based on the slot identification information, transports the incubator 1 to the turntable 65 by the transport means 6, and returns to steps S13 and S15. The cell state is measured. When it is determined in step S15 that the cells have reached confluence from the state of the cells, the controller 21 determines the force of the cells reaching the required amount based on the number of cells or the number of incubators obtained by measuring the cell state. It is determined whether or not it is (step S21). If it is determined in step S21 that the cells are not necessary, the incubator 1 in which no cells have been seeded is extracted from the slot 91 of the culture unit 39 based on the slot identification information, and transported to the turntable 65. Then, subculture is performed from the primary culture incubator 1 using the pipetting mechanism that constitutes the cell seeding means and the cell seeding pipenose 67i of the imaging unit 67. And passage (step S23).

After step S23, the controller 21 creates related identification information for identifying the incubator 1 of each subculture based on the identification information of the incubator 1 of the primary culture, and uses the barcode printing / pasting device 89 The label on which the barcode corresponding to the created discrimination information is printed is attached to the subculture incubator 1 (step S25). After step S25, the incubator 1 for primary culture and the incubator 1 for subculture are transported to and accommodated in the slot 91 of the culture unit 39, and the culture in step S17 is performed. At this time, the controller 21 stores the related identification information of the subculture incubator 1 stored in the slot 91 in association with the slot identification information of the slot 91. If no abnormality is detected in step S13, step S17, The subculture cycle of step S13, step S15, step S21 to step S25, and step 17 is repeated to increase the number of cells while subculturing.

 The control unit 21 repeats the subculture cycle, and when it is determined in step S21 that the cells have the required amount, the transport unit 6 sequentially transports the incubator 1 in the cell culture device to the shutter 43 of the opening / closing unit 19. Then, the shutter 43 is opened and carried out of the apparatus (step S27). When the unloading of the incubator 1 is completed, the control section 21 closes the shutter 43 of the next opening / closing section 19 and turns on the ultraviolet lamp 82 (step S29).

 On the other hand, the controller 21 repeats the subculture cycle, and if it is determined in step S13 that an abnormality such as cell death or concomitant generation has occurred, the controller 21 turns on the alarm function, turns on the alarm lamp 113, and turns on the alarm lamp 113. An alarm sound is emitted from the alarm speaker 115 (step S31). The operator who receives the alarm checks the state of the abnormality that has occurred on the monitor of the controller 21 and the like.The original incubator in which cells were subcultured to the incubator 1 in which the abnormality occurred and the incubator 1 in which the abnormality occurred If it is determined that the cultivation of the incubator 1 in which the cells have been subcultured from the incubator 1, i.e., the incubator 1 related to the incubator 1 in which the abnormality has occurred, should be stopped, the Turn off the stop command switch 1Γ7. After step S31, the controller 21 detects the state of the culture stop command switch 1Γ7 (step S33), and when the stop command switch 117 is turned on, the identification information or the related identification information of the incubator 1 in which the abnormality has occurred. The incubator 1 having the identification information or the related identification information associated with the incubator 1 in which the abnormality has occurred is extracted based on the (Step S35), and the incubator 1 in which the abnormality has occurred in Step S27 is taken out of the apparatus and Then, the corresponding other incubator 1 extracted in step S35 is also carried out of the apparatus. At this time, the controller 21 associates the identification information given to the incubator 1 or related identification information with the slot identification information of the slot 91 in which the incubator 1 is accommodated, as described above, and Manages incubator 1 in body 17. Therefore, the extraction of the incubator 1 in step S35 is also performed based on the slot identification information.

As described above, in the cell culture apparatus of the present embodiment, the culture means 15, the medium exchange means 9, the cell state measuring means 5, the cell seeding means 11, and the transport means 6 are housed in a hermetically formed casing. . Then, the cell seeding means uses the cell state measuring means to predict the cells in the incubator. When it is determined that the cell is in the set state, the cells in the incubator that measured the state of the cells are seeded in another incubator in which no cells have been seeded. Therefore, culturing cells, changing the culture medium in the incubator, measuring the state of the cells in the incubator, re-inoculating the cells in the incubator with the confluent cell state in the incubator into new medium, The work involved in the subculture can be performed in one device without manual intervention. In addition, it is necessary to suppress the generation of contamination during the work involved in these subcultures. All the work involved in these subcultures is performed in a hermetically sealed housing in a state separated from the external environment. And the occurrence of contamination can be suppressed. Therefore, there is no need for a skilled worker to perform the complicated work associated with the subculture manually, and the workability of the subculture can be improved.

 Furthermore, since the workability of subculture can be improved, subculture can be easily performed. In addition, it will be possible to mass-produce useful components produced from cells cultured by subculture or tissues for regenerative medicine.

 Furthermore, in the cell culture apparatus of the present embodiment, the cell state measuring means 5, the medium exchange means 9, and the cell seeding means 11 are integrally formed by a pitting mechanism, an imaging unit 67, a turntable 65, and the like. The device can be miniaturized. In addition, the culturing unit 39 constituting the culturing means 15 accommodates the incubator 1 one by one and has a plurality of slots 91 separated by a partition wall 93, so that the culturing unit accommodated in the adjacent slots 91 is provided. This prevents contact between the incubator 1 and contamination and cross-contamination due to microbial infection between the incubators.

 Furthermore, in the cell culture device of the present embodiment, the housing 17 is separated from the medium exchange and cell seeding unit 37 and the culture unit 39 by the internal partition wall 23, so that the internal environment of the culture unit is reduced. Adjustment can be made easily. However, as shown in FIG. 6, the inside of the casing 17 is separated by the internal partition 23 into a medium exchange and cell seeding section 37 and a culture section 39, that is, a configuration in which the culture space of the culture means 15 is not separated from the other space. You can also.

In addition, the cell state measuring means 5 has a microscope CCD camera unit 67g for photographing the incubator 1, and analyzes an image photographed by the microscope CCD camera unit 67g to measure the state of the cells. This eliminates the need for the operator to measure the state of the cells by observation with a microscope or the like, thereby improving workability of subculture. I can do it.

 Further, in the cell culture apparatus of the present embodiment, the ID reading means including the barcode reader 63 for reading the barcode printed on the label attached to the culture vessel 1 carried in from the opening / closing section 19 of the housing 17. Has three. For this reason, information on the type and origin of cells contained in the incubator 1 for primary culture carried into the device, and information on the state of the cells in the incubator 1 and the position of the incubator 1 in the device Information such as information on the incubator 1 can be easily managed. In addition, by searching and calling a preset culture protocol based on the identification information, the workability of subculture can be further improved.

Further, in the cell culture device of the present embodiment, the incubator 1 containing the medium in which the cells are inoculated by the cell inoculating means 11 is placed in the incubator 1 in which the cells to be inoculated in the incubator 1 are collected. A related identification information assigning means 13 is provided for creating and attaching related identification information that is related to the attached identification information. Then, the related identification information providing means 13 affixes a label printed with a barcode corresponding to the related identification information created by the percode printing / pasting device _{89 to} the incubator 1 for subculture. With such a configuration, therefore, information such as the origin, culture status, and position of the incubator in which cells have been replated for subculture can also be managed.

Further, in the cell culture device of the present embodiment, the controller 21 controls the incubator 1 for stopping the culturing in response to the stop command by the stop command switch 117 for commanding the stop of the culture of the specific incubator 1 to the transporting means 6. The incubator 1 is transported to the opening / closing section 19 of the casing 17 and discharged from the opening / closing section 19, and the culture is stopped from the incubator 1 other than the incubator 1 in which the culture is stopped according to the stop command in the casing 17. The incubator 1 to which the identification information or related identification information related to the identification information 1 or the related identification information 1 is attached is extracted and transported to the opening / closing section 19 of the housing 17 by the transporting means 6 and from the opening / closing section 19. Discharge. In addition, the controller 21 has a function of automatically performing the same operation as described above when the cell state measuring means 5 detects an abnormality in culture. Therefore, when an abnormality is found in the incubator, for example, when cell death or virus infection is found, the incubator in which the original cells inoculated in the incubator in which the abnormality was found are cultured The incubator, in which cells are replated and passaged from the incubator in which an abnormality is found, can be carried out of the apparatus. Furthermore, in the cell culture device of the present embodiment, the controller 21 closes the opening / closing portion 19 of the housing 17 when the incubator 1 containing the medium in which the cells are seeded is present in the housing 17. Hold to prevent another incubator 1 from being carried in. Therefore, if one cell culture device is used for culturing one cell to prevent cross-contamination, it is necessary to prevent an incubator containing another cell from being accidentally brought into the device. Can be.

 In addition, when the controller 21 inputs or reads the identification information attached to the incubator 1 before carrying it into the housing 17, the incubator 1 containing the medium in which the cells are seeded in the housing 17 is present. When the identification information input or read and the identification information attached to the incubator 1 in the housing 17 are used, the identification information to be carried in and used in the incubator 1 is read. When it is detected that the cells have different origins from the cells in the incubator 1 in the housing 17, the carrying-in of the incubator 1 is prevented by keeping the opening / closing portion 19 of the housing 7 closed. Therefore, if one cell culture device is used for culturing one cell to prevent cross-contamination, it is necessary to prevent incubators containing cells of different origins from being accidentally loaded into the device. Can be.

 Further, the cell culture apparatus of the present embodiment has an air nozzle 103 and an air supply line 105 which constitute air pressure adjusting means for adjusting the air pressure in the housing 7 to a more positive pressure than the air pressure around the housing 17. ing. For this reason, the opening / closing section 19 of the housing 17 does not need to have a structure such as an air-lock type that makes it difficult for the atmosphere outside the housing 17 to flow into the housing 17. Can be simplified. However, the air pressure adjusting means may not be provided, and the opening / closing portion 19 of the housing 17 may be configured such that the atmosphere outside the housing 17 does not easily flow into the housing # 7, for example, an air lock type.

As described above, the casing 7 is separated from the medium exchange and cell seeding section 37 and the culture section 39 by the internal partition 23, but the medium exchange and cell seeding section 37 is a pipe for the incubator. The risk of causing contamination is higher than that of the culture unit 39 due to access from the site. Therefore, if the pressure of the culture unit 39 is higher than that of the medium exchange and cell seeding unit 37, even if the culture unit 39 becomes contaminated, the inflow of contaminated air into the culture unit 39 can be prevented, which is preferable. . This can be easily realized by controlling the opening and closing of an electromagnetic valve for adjusting the amount of air flow from the air nozzle 103.

In each of the drawings referred to in the present embodiment, a force indicating a petri dish-shaped incubator is shown. Incubators having various shapes and structures can be used. Further, in the cell culture apparatus of the present embodiment, the space in which the cell seeding means 11, the medium exchange means 9, and the like constituting the cell seeding means 11, the medium exchange means 9, etc., and the imaging unit 67 and the turntable 65 are installed are provided. It has an ultraviolet lamp 82 for irradiating rays and a controller 21 for controlling lighting of ultraviolet rays. Then, the controller 21 stops the irradiation of the ultraviolet rays by the ultraviolet lamp 82 when the incubator is on the turntable 65. Therefore, microorganisms in the space where the culture medium in the incubator 1 is exposed to the air in the housing 17 can be killed without affecting the culture.

 Further, in the cell culture apparatus of the present embodiment, the ID reading means 3 is provided inside the housing 17, but as shown in FIGS. 7 and 8, the ID reading means 127 is provided outside the housing 17. You can also In this case, when culturing, after reading a bar code or the like printed on the label of the incubator 1 by the ID reading means 127, the incubator 1 is put into the housing 17 through the cell entrance 19a. Further, when the controller 21 has a function of preventing the incubator from being loaded with cells other than cells of the same origin as the incubator inhibiting means, the ID reading means is provided outside the housing 17. With the configuration provided with 127, it is not necessary to input the identification information with the input devices of the controller 21.

 In addition, the cell culture device of the present invention is not limited to the configuration of the present embodiment, and accommodates a culture unit, a medium exchange unit, a cell state measurement unit, a cell seeding unit, a transportation unit, and the like in an airtightly formed housing. With such a configuration, each means can be configured with various mechanisms and structures.

 As described above, according to the present invention, the workability of subculture can be further improved while preventing contamination / cross contamination.

Claims

A culturing means for culturing cells by accommodating at least one incubator containing a medium in which cells can be seeded and whose internal environment is adjustable;

 Medium exchange means for exchanging the medium in the incubator,

 Cell state measuring means for measuring the state of the cells in the incubator,

 Contract

 A cell-seeding seeding means for seeding the cells in the incubator cultured by the culture means in another incubator in which the cells have not been seeded;

 A hermetically sealable six housing that houses the culture means, the medium exchange means, the cell state measurement means, and the cell seeding means, and has at least one opening / closing part;

 The opening / closing section of the housing, at least the culture means, and the medium exchange means

A cell culture device, comprising: and a culture vessel transport means for transporting the culture vessel between the cell state measurement means and one of the cell seeding means.

 2. The cell culture device according to claim 1, wherein the culture means has a plurality of incubators accommodated one by one, and has a plurality of slots separated by partition walls. The housing has a space for accommodating the incubator constituting the culture means in the housing, and an internal partition separating a space other than the space for accommodating the incubator. 2. The cell culture device according to claim 1, wherein the cell culture device has at least one inner opening / closing portion.

 The housing has a space for accommodating the incubator constituting the culture means in the housing, and an internal partition separating a space other than the space for accommodating the incubator. 3. The cell culture device according to claim 2, wherein the cell culture device has at least one inner opening / closing portion.

The cell seeding means is characterized by disseminating the cells in the incubator whose cell state has been measured in another incubator in which no cells have been seeded, according to the measurement result by the cell state measuring means. 2. The cell culture device according to claim 1, wherein The cell seeding means is configured to perform another culture in which the cells in the incubator whose cell state has been measured are not seeded according to the measurement result of the cell state measuring means. 3. The cell culture device according to claim 2, wherein the cell culture device is seeded in a vessel. 7. The cell state measuring means according to claim 1, wherein the cell state measuring means has a camera for photographing the incubator, and analyzes an image photographed by the camera to measure a cell state. Cell culture equipment.

8. The cell state measuring means according to claim 2, wherein the cell state measuring means has a camera for photographing the incubator, and analyzes an image photographed by the camera to measure a cell state. Cell culture equipment.

 9. The cell culture device according to claim 1, further comprising identification information reading means for reading identification information attached to the incubator before or after being carried into the casing. .

 10. The cell culture device according to claim 2, further comprising identification information reading means for reading the identification information attached to the incubator either before or after carrying into the casing.

 11. The method according to claim 1, further comprising: identification information reading means for reading identification information attached to the incubator either before or after loading into the casing by the cell seeding means. Cell culture equipment.

 12. The method according to claim 2, further comprising identification information reading means for reading the identification information attached to the incubator either before or after the cell is seeded into the casing by the cell seeding means. Cell culture equipment.

13. A related identification information providing means for providing the related identification information created in association with the identification information attached to the incubator from which the cells were collected to the incubator in which the cells were inoculated by the cell seeding means. The cell culture device according to claim 1, further comprising:

 14. A related identification information providing means for providing the related identification information created in association with the identification information attached to the incubator from which the cells were collected to the incubator in which the cells were seeded by the cell seeding means. 3. The cell culture device according to claim 2, further comprising:

15. Include a stop command switch to command the suspension of cultivation of a specific incubator, and incubate the incubator for suspending the cultivation in response to the suspension instruction by the above-mentioned transport means. A culture in which the culture is transported to the opening / closing section, discharged from the opening / closing section, and cultivated in response to the stop command from an incubator other than the incubator in which the culture is stopped in response to the stop instruction in the casing. The incubator to which the identification information or the identification information related to the container identification information or the related identification information is attached is extracted, transported to the opening / closing section of the housing by the transport means, and discharged therefrom. 14. The cell culture apparatus according to claim 13, wherein the cell culture means is provided.

 16. In response to the determination of the abnormality by the cell state measuring means, the culture device in which the abnormality is determined is transported to the opening / closing section of the housing by the transporting means, and is discharged from the opening / closing section. From the incubators other than the incubator in which the abnormality is determined in the housing, the incubator to which the identification information or the related identification information related to the identification information or the related identification information of the incubator in which the abnormality is determined is attached. A culture suspending means for extracting, transporting to the opening / closing section of the housing by the transporting means, and discharging from the opening / closing section

14. The apparatus for culturing cells according to claim 13, wherein the apparatus has _ stages.

 17. When there is an incubator containing a medium in which cells have been seeded in the housing, an alarm and / or holding the opening / closing part of the housing closed so that the incubator of another incubator 2. The Itoda spore cultivation apparatus according to claim 1, further comprising a culture vessel carry-in preventing means for preventing carry-in into the housing.

 18. From the identification information recognized by inputting or reading the identification information attached to the incubator before loading into the above-mentioned housing, the cells in the incubator with this identification information are present in the above-mentioned housing. When detecting that the origin is different from the cell in the incubator, at least one of an alarm and keeping the opening / closing part of the housing closed is used for the incubator containing cells of different origin. 2. The cell culture device according to claim 1, further comprising a culture vessel loading prevention means for preventing loading into the housing.

19. The Itoda spore culturing apparatus according to claim 1, further comprising an air pressure adjusting means for adjusting an air pressure in the housing to a positive pressure than an air pressure around the housing.

20. An ultraviolet irradiation unit for irradiating at least a space in the casing where cells are seeded by the cell seeding unit and a space where a medium is exchanged by the medium exchange unit with ultraviolet irradiation, wherein the ultraviolet irradiation unit is The above cell seeding hands 2. The cell according to claim 1, wherein irradiation of ultraviolet rays is stopped when a culture vessel is present in at least one of a space in which the cells are seeded by the step and a space in which the medium is exchanged by the medium exchange means. Culture equipment.

 21. The space according to claim 3, wherein the pressure in a space other than the space accommodating the incubator is lower than the space accommodating the incubator constituting the culture means in the housing. Cell culture equipment. .

22. The space according to claim ^{4, wherein} the pressure in a space other than the space accommodating the incubator is lower than the space accommodating the incubator constituting the culture means in the housing. Cell culture device