Patent Publication Number: US-2018044624-A1

Title: Cell Culture Device, Cartridge for Culture Medium Replacement Use, and Method for Replacing Culture Medium

Description:
TECHNICAL FIELD 
     The present disclosure relates to a cell culture device for culturing cells. Furthermore, the present disclosure relates to a cartridge for culture medium replacement use utilized in a cell culture device. Moreover, the present disclosure relates to a method for replacing a culture medium using a cartridge for culture medium replacement use. 
     BACKGROUND 
     In the related art, there is known a culture device used for culturing cells. In such a culture device, a culture medium replacement mechanism for periodically replacing a liquid culture medium inside a culture container is usually installed in order to prevent gradual deterioration of an internal culture environment of the culture container. As an example of the culture medium replacement mechanism, for example, Patent Document 1 is disclosed. In Patent Document 1, there is disclosed a mechanism in which a culture container placed on a culture medium replacement stage (container mounting stand), a culture medium storage container and a collection container are connected by different tubes, a new culture medium is supplied to the culture container from the culture medium storage container via one tube, and an old culture medium is collected from the culture container to the collection container via the other tube. 
     PRIOR ART DOCUMENTS 
     Patent Documents 
     Patent Document 1: Japanese laid-open publication No. 2008-271850 
     However, in the culture medium replacement mechanism provided in the related art, other devices such as a culture medium storage container, a collection container, a culture medium temperature regulator and a culture medium time adjustor are concentrated around the culture medium replacement stage. Therefore, due to physical limitations, a liquid flow path connecting the culture container and the culture medium storage container and the like becomes redundant. In addition, protein gradually adheres to an inner wall of the liquid flow path when in use, thereby contaminating the liquid flow path. Thus, it is necessary to periodically replace the liquid flow path. Since the liquid flow path is a tube, it is difficult to automate the replacement. It is also difficult to ensure the workability due to the concentration of devices. 
     Furthermore, in the culture media replacement mechanism provided in the related art, it is not possible to start a treatment on a subsequent culture container until a series of treatments such as culture medium supply, culture medium replacement, culture medium collection and flow path cleaning are completed for one culture container. Thus, the operation time in the case of treating a plurality of culture containers is prolonged. 
     The present disclosure provides some embodiments of a cell culture device, a cartridge for culture medium replacement use and a method for replacing a culture medium, which are capable of shortening and automatically replacing a liquid flow path. 
     SUMMARY 
     According to one embodiment of the present disclosure, there is provided a cell culture device, including: an incubator part configured to accommodate a closed-system culture container; a cartridge for culture medium replacement use having a liquid supply flow path and a liquid collection flow path, the cartridge removably attachable to the culture container; a culture medium supply part configured to supply a liquid culture medium to the liquid supply flow path of the cartridge for culture medium replacement use; a culture medium replacement part configured to cause the liquid culture medium existing in the liquid supply flow path to flow into the culture container and to cause the liquid culture medium existing in the culture container to flow out to the liquid collection flow path in a state in which the cartridge for culture medium replacement use is connected to the culture container; and a culture medium collection part configured to collect the liquid culture medium from the liquid collection flow path of the cartridge for culture medium replacement use, wherein the cartridge for culture medium replacement use is movable between the culture medium supply part, the culture medium replacement part and the culture medium collection part. 
     The device according to the present disclosure may further include: a cartridge conveying part configured to move the cartridge for culture medium replacement use between the culture medium supply part, the culture medium replacement part and the culture medium collection part. 
     The device according to the present disclosure may further include: a flow path cleaning part configured to clean the liquid supply flow path and the liquid collection flow path of the cartridge for culture medium replacement use, wherein the cartridge for culture medium replacement use may move between the culture medium supply part, the culture medium replacement part, the culture medium collection part and the flow path cleaning part. 
     In the device according to the present disclosure, the culture medium replacement part may be installed adjacent to the incubator part, and a culture medium replacement in the culture container may be performed via the cartridge for culture medium replacement use while keeping the culture container accommodated in the incubator part. 
     In the device according to the present disclosure, a culture medium analysis part configured to analyze a collected liquid culture medium may be installed in the culture medium collection part. 
     The device according to the present disclosure may further include: a cartridge storage part configured to store a plurality of unused cartridges for culture medium replacement use; and a cartridge collection part configured to collect a used cartridge for culture medium replacement use. 
     In the device according to the present disclosure, the liquid supply flow path may include a first inflow port connectable to the culture medium supply part, a first outflow port connectable to an inflow port of the culture container, a liquid storage chamber configured to bring the first inflow port and the first outflow port into communication with each other and a ventilation port communicating with the liquid storage chamber, and the liquid collection flow path may include a second inflow port connectable to an outflow port of the culture container, a second outflow port connectable to the culture medium collection part and a curved flow path configured to bring the second inflow port and the second outflow port into communication with each other, the curved flow path including a plurality of bent portions or curved portions. 
     In the device according to the present disclosure, the curved flow path may have a serpentine shape or a spiral shape. 
     In the device according to the present disclosure, the first outflow port and the second inflow port may be formed adjacent to each other. 
     A cartridge for culture medium replacement use according to the present disclosure, including: a liquid supply flow path; and a liquid collection flow path, wherein the cartridge is removably attachable to a closed-system culture container and is movable between a culture medium supply part, a culture medium replacement part and a culture medium collection part of a cell culture device. 
     In the cartridge according to the present disclosure, the liquid supply flow path may include a first inflow port connectable to the culture medium supply part, a first outflow port connectable to an inflow port of the culture container, a liquid storage chamber configured to bring the first inflow port and the first outflow port into communication with each other and a ventilation port communicating with the liquid storage chamber, and the liquid collection flow path may include a second inflow port connectable to an outflow port of the culture container, a second outflow port connectable to the culture medium collection part and a curved flow path configured to bring the second inflow port and the second outflow port into communication with each other, the curved flow path including a plurality of bent portions or curved portions. 
     In the cartridge according to the present disclosure, the curved flow path may have a serpentine shape or a spiral shape. 
     In the cartridge according to the present disclosure, the first outflow port and the second inflow port may be formed adjacent to each other. 
     A method for replacing a culture medium according to the present disclosure, including: a culture medium supply step of connecting a cartridge for culture medium replacement use having a liquid supply flow path and a liquid collection flow path to a culture medium supply part, supplying a liquid culture medium from the culture medium supply part to the liquid supply flow path, and separating the cartridge for culture medium replacement use from the culture medium supply part; a culture medium replacement step of connecting the cartridge for culture medium replacement use to a closed-system culture container, causing the liquid culture medium existing in the liquid supply flow path to flow into the culture container, causing the liquid culture medium existing in the culture container to flow out to the liquid collection flow path, and separating the cartridge for culture medium replacement use from the culture container; and a culture medium collection step of connecting the cartridge for culture medium replacement use to a culture medium collection part, collecting the liquid culture medium from the liquid collection flow path to the culture medium collection part, and separating the cartridge for culture medium replacement use from the culture medium collection part. 
     The method according to the present disclosure may further include a cleaning step of cleaning the liquid supply flow path and the liquid collection flow path of the cartridge for culture medium replacement use after the culture medium collection step. 
     In the method according to the present disclosure, the culture medium replacement step may be performed in a state in which the culture container is accommodated in an incubator part. 
     The method according to the present disclosure may further include a culture medium analysis step of analyzing a collected liquid culture medium after the culture medium collection step. 
     According to the present disclosure, a cartridge for culture medium replacement use capable of accommodating a predetermined amount of liquid culture medium moves between a culture medium supply part, a culture medium replacement part and a culture medium collection part. This makes it possible to perform a culture medium replacement in a state in which a culture container is separated from the culture medium supply part and the culture medium collection part. Therefore, it is possible to shorten a liquid flow path. In addition, the liquid flow path is formed in the cartridge for culture medium replacement use, namely a cartridge-type part. This makes it easy to automate the replacement of the liquid flow path. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a schematic top plan view showing a configuration of an automatic culture system according to an embodiment of the present disclosure. 
         FIG. 2  is a control block diagram of an automatic culture system according to an embodiment of the present disclosure. 
         FIG. 3  is a front view showing a container conveying part used in an embodiment of the present disclosure. 
         FIG. 4  is a schematic plan view showing a configuration in the vicinity of a liquid storage supply part and an incubator part used in an embodiment of the present disclosure. 
         FIG. 5A  is a schematic view showing a configuration of a cartridge for culture medium replacement use utilized in an embodiment of the present disclosure. 
         FIG. 5B  is a schematic view showing a configuration of a first modification of the cartridge for culture medium replacement use. 
         FIG. 5C  is a schematic view showing a configuration of a second modification of the cartridge for culture medium replacement use. 
         FIG. 6A  is a schematic plan view showing a structure of a valve of the cartridge for culture medium replacement use shown in  FIG. 5A . 
         FIG. 6B  is a sectional view taken along line A-A of the valve shown in  FIG. 6A . 
         FIG. 7A  is a view corresponding to  FIG. 6B  and explaining an operation of the valve. 
         FIG. 7B  is a view corresponding to  FIG. 6B  and explaining the operation of the valve. 
         FIG. 7C  is a view corresponding to  FIG. 6B  and explaining the operation of the valve. 
         FIG. 8A  is a view corresponding to  FIG. 5A  and explaining a culture medium supply step. 
         FIG. 8B  is a view corresponding to  FIG. 5A  and explaining the culture medium supply step. 
         FIG. 9A  is a view corresponding to  FIG. 5A  and explaining a culture medium replacement step. 
         FIG. 9B  is a view corresponding to  FIG. 5A  and explaining the culture medium replacement step. 
         FIG. 10A  is a view corresponding to  FIG. 5A  and explaining a culture medium collection step. 
         FIG. 10B  is a view corresponding to  FIG. 5A  and explaining the culture medium collection step. 
         FIG. 11A  is a view corresponding to  FIG. 5A  and explaining a flow path cleaning step. 
         FIG. 11B  is a view corresponding to  FIG. 5A  and explaining the flow path cleaning step. 
         FIG. 11C  is a view corresponding to  FIG. 5A  and explaining the flow path cleaning step. 
         FIG. 11D  is a view corresponding to  FIG. 5A  and explaining the flow path cleaning step. 
         FIG. 12A  is an internal top view showing a configuration of a first modification of a transport container. 
         FIG. 12B  is an internal side view showing the configuration of the first modification of the transport container. 
         FIG. 13  is a view corresponding to  FIG. 12B  and explaining an operation of the transport container. 
         FIG. 14  is an internal top view corresponding to  FIG. 12A  and showing a configuration of a second modification of the transport container. 
         FIG. 15A  is a schematic plan view showing a configuration of a modification example of the cell culture device. 
         FIG. 15B  is a sectional view taken along line B-B of the cell culture device in  FIG. 15A . 
         FIG. 15C  is a sectional view taken along line C-C of the cell culture device in  FIG. 15A . 
         FIG. 15D  is a left side view of the cell culture device shown in  FIG. 15A . 
     
    
    
     DETAILED DESCRIPTION 
     Embodiments of the present disclosure will now be described in detail with reference to the accompanying drawings. Throughout the drawings attached hereto, for the convenience of illustration and ease of understanding, the scales, the aspect ratios and the like are changed and exaggerated from actual embodiments. 
     The culture device according to the present embodiment may be used for culturing all kinds of cells and may be used when culturing various cells including pluripotent stem cells such as (human) iPS cells, (human) ES cells or the like, chondrocytes such as bone marrow stromal cells (MSCs) or the like, dendritic cells, and so forth. In the present embodiment, hereinafter, an automatic culture system for automatically culturing iPS cells will be described. However, it should be noted that this is nothing more than an example. In other words, it should be noted that the culture device according to the present embodiment may be used even if cells are not automatically cultured as in the present embodiment. 
     &lt;Overall Configuration&gt; 
     First, the configuration of the automatic culture system according to the present embodiment will be described. 
     As shown in  FIG. 1 , the automatic culture system according to the present embodiment includes a raw material storage device  10  configured to store raw material cells, a container transfer part  60  configured to transfer a transport container  70  (see  FIG. 3 ) in which cells or the like are accommodated in a sealed state, and cell culture devices  20  and  30  configured to receive the transport container  70  transferred by the container transfer part  60 , takes out a closed-system culture container  75  in which cells are accommodated from the transport container  70 , and cultures the cells contained in the taken-out culture container  75 . 
     In the present embodiment, the mode using iPS cells as mentioned above will be described. Thus, the raw material storage device  10  includes an iPS cell establishing device  11  configured to establish iPS cells. In addition, the raw material storage device  10  includes a unit thermostat bath, a centrifugal separator, an automatic blood cell counter, an automatic magnetic cell separator, a flow cytometer, a gene introduction device, and the like. 
     The cell culture devices  20 ,  30  of the present embodiment include a plurality of (four, in the embodiment shown in  FIG. 1 ) iPS cell automatic culture devices  20  configured to automatically culture iPS cells, and a plurality of (eight, in the embodiment shown in  FIG. 1 ) differentiated cell automatic culture devices  30  configured to automatically culture differentiated cells differentiated from the iPS cells. In the present embodiment, when merely referred to as “cell culture device”, it means the iPS cell automatic culture device  20 , the differentiated cell automatic culture device  30 , or both the iPS cell automatic culture device  20  and the differentiated cell automatic culture device  30 . Unless specifically mentioned otherwise, when merely referred to as “cells” in the present embodiment, it means raw material cells such as somatic cells forming the basis of iPS cells or the like, iPS cells, differentiated cells, or two or all of the raw material cells, the iPS cells and the differentiated cells. 
     As shown in  FIG. 3 , the transport container  70  includes a plurality of (eight, in  FIG. 3 ) shelves  71  configured to support the plurality of culture containers  75 . Each of the culture containers  75  is placed on each of the shelves  71 . The transport container  70  is transferred by the container transfer part  60  in a state in which the plurality of culture containers  75  is accommodated in the transport container  70 . 
     As illustrated in  FIG. 2 , the iPS cell automatic culture device  20  includes a housing  22  illustrated in  FIG. 1 , a culture medium analysis part  24  configured to analyze liquid culture medium components that vary with the culture of iPS cells, a cell inspection removal part  25  configured to inspect the iPS cells and performs removal of the iPS cells having a bad state, a liquid storage supply part  26  configured to store and supply a liquid containing a liquid culture medium or a proteolytic enzyme and to perform a pre-treatment before iPS cells are seeded, to seed iPS cells or to collect iPS cells, an incubator part  27  configured to hold the culture container  75  and automatically adjust one or all of a temperature, a humidity and a gas concentration, and a discharge part  28  configured to discharge downward from the housing  22  a waste liquid containing a used liquid culture medium, a used cleaning liquid, a used reagent or the like, which is used in the iPS cell automatic culture device  20 . Furthermore, the iPS cell automatic culture device  20  includes an in-device transfer part  23  configured to transfer the culture container  75  or the like inside the iPS cell automatic culture device  20 . The liquid storage supply part  26  described above also has a function of inverting the culture container  75  to be upside down. Incidentally, in the case where the closed-system culture container  75  is used as in the present embodiment, it is not necessarily required to manage the internal humidity of the incubator part  27  at a strict level as compared with the case of using an open-system culture container. This makes it possible to simplify the management of a cell culture environment. By employing the closed-system culture container  75  of this type, there is no fear that contamination from the air occurs. Furthermore, the transfer becomes easy. 
     The liquid storage supply part  26  described above appropriately supplies a liquid culture medium from an inflow port (not shown) into the culture container  75 , thereby automatically replacing an old liquid culture medium existing in the culture container  75  with a new liquid culture medium. Based on the information of the iPS cells acquired, the cell inspection removal part  25  selectively peels off defective iPS cells from an ECM (Extracellular Matrix) coated on the surface of a film (not shown) of the culture container  75 . Thereafter, the liquid storage supply part  26  supplies a liquid culture medium from the inflow port into the culture container  75 , thereby pushing out floating defective iPS cells from the culture container  75  through an outflow port (not shown). As the method of selectively peeling off the iPS cells existing in the culture container  75 , it may be possible to use a method of irradiating ultrasonic waves or light onto the iPS cells or a method of applying a physical force from outside the culture container  75 . When using this method, it may be possible to use a proteolytic enzyme in combination. 
     Furthermore, the liquid storage supply part  26  appropriately supplies a proteolytic enzyme from the inflow port into the culture container  75 , thereby peeling off the iPS cells from the ECM coated on the surface of a film of the culture container  75 . Thereafter, the liquid storage supply part  26  supplies a liquid culture medium from the inflow port into the culture container  75 , whereby floating iPS cells are pushed out from the culture container  75  through the outflow port. The iPS cells thus pushed out are diluted into a suspension and are then accommodated (seeded) in a plurality of other culture containers  75 . In this way, the iPS cell automatic culture device  20  automatically performs the subculture of the iPS cells. 
     The internal temperature of the iPS cell automatic culture device  20  is adjusted by the incubator part  27  so that the internal temperature becomes, for example, about 37 degrees C. Furthermore, the gas concentration in the iPS cell automatic culture device  20  is adjusted by the incubator part  27  by appropriately adding carbon dioxide or nitrogen. If necessary, the humidity may be adjusted by the incubator part  37  so as to become about 100%. 
     As illustrated in  FIG. 2 , the differentiated cell automatic culture device  30  includes a housing  32  illustrated in  FIG. 1 , a culture medium analysis part  34  configured to analyze liquid culture medium components that vary with the culturing of differentiated cells, a cell inspection removal part  35  configured to inspect the differentiated cells and perform removal of the differentiated cells having a bad state, a liquid storage supply part  36  configured to store and supply a liquid containing a liquid culture medium or a proteolytic enzyme and to perform a pre-treatment before differentiated cells are seeded, seed differentiated cells or collect differentiated cells, an incubator part  37  configured to hold the culture container  75  and automatically adjust one or all of a temperature, a humidity and a gas concentration, and a discharge part  38  configured to discharge downward from the housing  32  a waste liquid containing a used liquid culture medium, a used cleaning liquid, a used reagent or the like, which is used in the differentiated cell automatic culture device  30 . Furthermore, the differentiated cell automatic culture device  30  includes an in-device transfer part  33  configured to transfer the culture container  75  or the like within the differentiated cell automatic culture device  30 . 
     The liquid storage supply part  36  described above appropriately supplies a liquid culture medium from the inflow port into the culture container  75 , thereby automatically replacing an old liquid culture medium existing in the culture container  75  with a new liquid culture medium. Based on the information of the differentiated cells acquired, the cell inspection removal part  35  selectively peels off defective differentiated cells from the ECM coated on the surface of a film  77  of the culture container  75 . Thereafter, the liquid storage supply part  36  supplies a liquid culture medium from the inflow port into the culture container  75 , thereby pushing out floating defective differentiated cells from the culture container  75  through the outflow port. As the method of selectively peeling off the differentiated cells existing in the culture container  75 , it may be possible to use a method of irradiating ultrasonic waves or light onto the differentiated cells or a method of applying a physical force from outside the culture container  75 . When using this method, it may be possible to use a proteolytic enzyme in combination. 
     Furthermore, the liquid storage supply part  36  appropriately supplies a proteolytic enzyme from the inflow port into the culture container  75 , thereby peeling off the differentiated cells from the ECM coated on the surface of the film of the culture container  75 . Thereafter, the liquid storage supply part  36  supplies a liquid culture medium from the inflow port into the culture container  75 , whereby floating differentiated cells are pushed out from the culture container  75  through the outflow port. The differentiated cells thus pushed out are diluted into a suspension and are then accommodated (seeded) within a plurality of other culture containers  75 . In this way, the differentiated cell automatic culture device  30  automatically performs the subculture of the differentiated cells. 
     The internal temperature of the differentiated cell automatic culture device  30  is adjusted by the incubator part  37  so that the internal temperature becomes, for example, about 37 degrees C. Furthermore, the gas concentration within the differentiated cell automatic culture device  30  is adjusted by the incubator part  37  by appropriately adding carbon dioxide or nitrogen. When inducing differentiation, the liquid storage supply part  36  of the differentiated cell automatic culture device  30  may supply a liquid culture medium including a differentiation-inducing factor. If necessary, the humidity may be adjusted by the incubator part  37  so as to become about 100%. 
     As illustrated in  FIG. 2 , the iPS cell automatic culture device  20  includes a control part  29  which is connected to each of the culture medium analysis part  24 , the cell inspection removal part  25 , the liquid storage supply part  26 , the incubator part  27 , the discharge part  28  and the in-device transfer part  23  so as to make communication therewith and which is configured to control them. The control part  29  has a function of, with respect to the iPS cell automatic culture device  20 , managing the status, managing the log, managing the culture schedule, or serving as a user interface. Furthermore, the differentiated cell automatic culture device  30  includes a control part  39  which is connected to each of the culture medium analysis part  34 , the cell inspection removal part  35 , the liquid storage supply part  36 , the incubator part  37 , the discharge part  38  and the in-device transfer part  33  so as to make communication therewith and which is configured to control them. The control part  29  has a function of, with respect to the differentiated cell automatic culture devices  30 , managing the status, managing the log, managing the culture schedule, or serving as a user interface. 
     The iPS cell establishing device  11  is similar in configuration to the iPS cell automatic culture device  20  and the differentiated cell automatic culture device  30 . That is to say, as illustrated in  FIG. 2 , the iPS cell establishing device  11  includes a housing  12   b  illustrated in  FIG. 1 , a culture medium analysis part  14  configured to analyze a liquid culture medium, a cell inspection removal part  15  configured to inspect the raw material cells and perform removal of the raw material cells having a bad state, a liquid storage supply part  16  configured to store and supply a liquid containing a liquid culture medium or a proteolytic enzyme, an incubator part  17  configured to automatically adjust one or all of a temperature, a humidity and a gas concentration in the housing  12   b , and a discharge part  18  configured to discharge downward from the housing  12   b  a waste liquid containing a used liquid culture medium, a used cleaning liquid, a used reagent or the like, which is used in the iPS cell establishing device  11 . Furthermore, the iPS cell establishing device  11  further includes an in-device transfer part  13  configured to transfer the culture container  75  within the iPS cell establishing device  11 . Moreover, the iPS cell establishing device  11  includes a control part  19  which is connected to each of the culture medium analysis part  14 , the cell inspection removal part  15 , the liquid storage supply part  16 , the incubator part  17 , the discharge part  18  and the in-device transfer part  13  so as to make communication therewith and which is configured to control them. Each of the control parts  19 ,  29  and  39  is connected to an external device  90  such as, e.g., a personal computer or the like. 
     As illustrated in  FIG. 3 , the container transfer part  60  of the present embodiment includes a holding part  61  which holds the transport container  70  while suspending downward the transport container  70 . The container transfer part  60  is configured to move along a rail  65  installed in a ceiling. 
     As illustrated in  FIG. 1 , the iPS cell automatic culture device  20  includes a loading part  21  configured to load the culture container  75  from the transport container  70  therethrough. In some embodiments, the loading part  21  may include a cell loading/unloading part (not shown) for loading the iPS cells accommodated in the culture container  75  therethrough and for unloading the cultured iPS cells therefrom, and a material loading part (not shown) for loading materials accommodated in other airtight containers therethrough. Similarly, as illustrated in  FIG. 1 , the differentiated cell automatic culture device  30  includes a loading part  31  configured to load the culture container  75  from the transport container  70  therethrough. In some embodiments, the loading part  31  may include a cell loading/unloading part (not shown) for loading the iPS cells accommodated in the culture container  75  therethrough and for unloading the cultured differentiated cells therefrom, and a material loading part (not shown) for loading materials accommodated in other airtight containers therethrough. In the present embodiment, the materials may include a liquid culture medium, a reagent, a cleaning liquid, a culture plate, a vial, a filter, a needle, and so forth. Furthermore, as illustrated in  FIG. 1 , the iPS cell establishing device  11  includes a loading part  12   a  configured to load the transport container  70  therethrough. 
     As illustrated in  FIG. 1 , the automatic culture system of the present embodiment includes a sterilizing device  1  for sterilizing the interior of the transport container  70 , an iPS cell analysis device  80  which receives the iPS cells cultured in the iPS cell automatic culture device  20  from the loading part  81  at a predetermined timing and which inspects the iPS cells thus received, a differentiated cell analysis device  85  which receives the differentiated cells cultured in the differentiated cell automatic culture devices  30  from the loading part  86  at a predetermined timing and which inspects the differentiated cells thus received, and a freezing storage device  40  which receives the iPS cells, the differentiated cells or both cultured in the automatic culture devices  20  and  30  from the loading part  41  and which freezes and stores the iPS cells, the differentiated cells or both thus received. A plurality of freezing storage devices  40  may be provided. The entire room may be cooled and may serve as a freezer. In the case where the plurality of freezing storage devices  40  are installed in the room or in the case where the room itself serves as a freezer, the rail  65  may be installed in the ceiling of the room so that the container transfer part  60  can move along the rail  65 . 
     One example of the sterilizing device  1  described above may include a sterilizing device which sterilizes the interior of the transport container  70  by supplying a sterilizing gas such as a hydrogen peroxide gas or a high-temperature gas into the transport container  70 . Another example of the sterilizing device  1  may include a sterilizing device which sterilizes the interior of the transport container  70  by irradiating, for example, y rays or ultraviolet rays from the outside while keeping the transport container  70  in a sealed state. In addition, before the transport container  70  is loaded from the outside, the interior of the transport container  70  may be sterilized using, for example, y rays or ultraviolet rays. There may be a case where the liquid culture medium or the like contains protein or the like which is broken by y rays or ultraviolet rays. In this case, it is desirable that sterilization is performed by a sterilizing gas such as a hydrogen peroxide gas, a high-temperature gas or the like. 
     &lt;Configuration Around Liquid Storage Supply Part and Incubator Part&gt; 
     Next, the configuration around the liquid storage supply part and the incubator part will be described. 
     In the following descriptions, one, two or all of the liquid storage supply part  16  of the iPS cell establishing device  11 , the liquid storage supply part  26  of the iPS cell automatic culture devices  20  and the liquid storage supply part  36  of the differentiated cell automatic culture devices  30  will be referred to as a “liquid storage supply part  110 .” In the present embodiment, the liquid storage supply part  16  of the iPS cell establishing device  11 , the liquid storage supply part  26  of the iPS cell automatic culture devices  20  and the liquid storage supply part  36  of the differentiated cell automatic culture devices  30  have the same configuration. 
     In the following descriptions, one, two or all of the incubator part  17  of the iPS cell establishing device  11 , the incubator part  27  of the iPS cell automatic culture device  20  and the incubator part  37  of the differentiated cell automatic culture device  30  will be referred to as an “incubator part  101 .” In the present embodiment, the incubator part  17  of the iPS cell establishing device  11 , the incubator part  27  of the iPS cell automatic culture device  20  and the incubator part  37  of the differentiated cell automatic culture device  30  have the same configuration. 
     In the following descriptions, one, two or all of the culture medium analysis part  14  of the iPS cell establishing device  11 , the culture medium analysis part  24  of the iPS cell automatic culture devices  20  and the culture medium analysis part  34  of the differentiated cell automatic culture devices  30  will be referred to as a “culture medium analysis part  106 .” In the present embodiment, the culture medium analysis part  14  of the iPS cell establishing device  11 , the culture medium analysis part  24  of the iPS cell automatic culture devices  20  and the culture medium analysis part  34  of the differentiated cell automatic culture devices  30  have the same configuration. 
       FIG. 4  is a schematic plan view showing a configuration around the liquid storage supply part  110  and the incubator part  101 . As shown in  FIG. 4 , the liquid storage supply part  110  includes a cartridge for culture medium replacement use  200  having a liquid supply flow path  201  and a liquid collection flow path  202  (see  FIG. 5A ) and detachably installed in the culture container  75 , a culture medium supply part  102  configured to supply a liquid culture medium to the liquid supply flow path  201  of the cartridge for culture medium replacement use  200 , a culture medium replacement part  103  configured to cause the liquid culture medium inside the liquid supply flow path  201  to flow into the culture container  75  and cause the culture medium inside the culture container  75  to flow out toward the liquid collection flow path  202  in a state in which the cartridge for culture medium replacement use  200  is connected to the culture container  75 , a culture medium collection part  104  configured to collect the liquid culture medium from the liquid collection flow path  202  of the cartridge for culture medium replacement use  200 , and a cartridge conveying part  109  configured to move the cartridge for culture medium replacement use  200  between the culture medium supply part  102  and the culture medium replacement part  103  and the culture medium collection part  104 . In  FIG. 4 , thin arrows indicate the direction of movement of the cartridge for culture medium replacement use  200 , and thick arrows indicate the direction of movement of the liquid culture medium. 
     Furthermore, in the present embodiment, a culture medium analysis part  106  configured to analyze the components of the collected liquid culture medium is installed in the culture medium collection part  104 . 
     Moreover, in the present embodiment, as shown in  FIG. 4 , there is installed a flow path cleaning part  105  configured to clean the liquid supply flow path  201  and the liquid collection flow path  202  of the cartridge for culture medium replacement use  200 . The cartridge conveying part  109  is adapted to move the cartridge for culture medium replacement use  200  between the culture medium supply part  102 , the culture medium replacement part  103 , the culture medium collection part  104  and the flow path cleaning part  105 . 
     Furthermore, in the present embodiment, as shown in  FIG. 4 , there are further installed a cartridge storage part  107  configured to store a plurality of unused cartridges for culture medium replacement use  200  and a cartridge collection part  108  configured to collect the used cartridges for culture medium replacement use  200 . The cartridge conveying part  109  is configured to periodically discharge the used cartridge for culture medium replacement use  200  to the cartridge collection part  108  and periodically take out the unused cartridge for culture medium replacement use  200  from the cartridge storage part  107   
     As the cartridge conveying part  109 , it may be possible to use, for example, a conveying robot which has a hand capable of gripping a cartridge-type component and which is well-known in the related art. 
     &lt;Configuration of Cartridge for Culture Medium Replacement Use&gt; 
     Next, a configuration of the cartridge for culture medium replacement use  200  will be described in detail with reference to  FIG. 5A . 
     In the present embodiment, as shown in  FIG. 5A , the liquid supply flow path  201  includes a first inflow port  211  connectable to the culture medium supply part  102 , a first outflow port  212  connectable to the inflow port of the culture container  75 , a liquid storage chamber  213  configured to bring the first inflow port  211  and the first outflow port  212  into communication with each other, and a ventilation port  214  communicating with the liquid storage chamber  213 . A filter  218  for preventing intrusion of extraneous matter from ambient air is attached to the ventilation port  214 . 
     The capacity of the liquid storage chamber  213  is about 1 to 1.5 times as large as the capacity of the culture container  75  and is preferably larger than the capacity of the culture container  75 . Specifically, the capacity of the liquid storage chamber  213  is, for example, 30 ml. As shown in  FIG. 5A , an openable/closable valve  215  is installed in a flow path formed between the first inflow port  211  and the liquid storage chamber  213 . An openable/closable valve  216  is installed in a flow path formed between the liquid storage chamber  213  and the first outflow port  212 . Further, an openable/closable valve  217  is installed in a flow path formed between the liquid storage chamber  213  and the ventilation port  214 . By closing the respective valves  215 ,  216  and  217 , the liquid storage chamber  213  is hermetically sealed. 
     On the other hand, the liquid collection flow path  202  includes a second inflow port  221  connectable to the outflow port of the culture container  75 , a second outflow port  222  connectable to the culture medium collection part  104 , and a curved flow path  223  configured to bring the second inflow port  221  and the second outflow port  222  into communication with each other. The curved flow path  223  includes a plurality of bent portions or curved portions. By including the bent portions or curved portions, the curved flow path  223  can be formed to have a small flow path diameter and a large flow path length. Thus, the old culture medium flowing out from the culture container  75  can be collected so as not to be mixed with the new culture medium in the flow path. 
     In the example shown in  FIG. 5A , the curved flow path  223  has a serpentine shape. However, as long as the curved flow path  223  can collect the old medium so as not to be mixed with the new culture medium in the flow path, it is not necessarily required for the curved flow path  223  to have a serpentine shape. For example, as shown in  FIG. 5B , the curved flow path  223  may have a spiral shape. 
     The capacity of the curved flow path  223  is about 1 to 1.5 times as large as the capacity of the culture container  75  and is preferably larger than the capacity of the culture container  75 . Specifically, the capacity of the curved flow path  223  is, for example, 30 ml. In order to suppress the generation of a turbulent flow in the curved flow path  223 , the diameter of the curved flow path  223  may be 4 mm or less. As shown in  FIG. 5A , an openable/closable valve  225  is installed in a flow path formed between the second inflow port  221  and the curved flow path  223 . An openable/closable valve  226  is installed in a flow path formed between the curved flow path  223  and the second outflow port  222 . By closing the respective valves  225  and  226 , the curved flow path  223  is hermetically sealed. 
     In the example shown in  FIGS. 5A and 5B , the entire curved flow path extending from the second inflow port  221  to the second outflow port  222  is formed as a narrow curved flow path  223 . However, as shown in  FIG. 5C , within a range where no mixing of liquids occurs between the old culture medium and the new culture medium (for example, within ½ of the collection capacity), a portion of the flow path extending from the second inflow port  221  to the second outflow port  222  may be formed as a tank-shaped flow path  224  and the remaining portion of the flow path may be formed as a narrow curved flow path  223 . Considering the conductance of the flow path from the second inflow port  221  to the second outflow port  222 , it is preferable that a portion of the flow path is formed as the tank-shaped flow path  224 . 
     The first outflow port  212  and the second inflow port  221  are formed adjacent to each other. Thus, the first outflow port  212  and the second inflow port  221  can be easily connected to the inflow port and the outflow port of the culture container  75 , respectively. 
     Hereinafter, one, two or more or all of the valve  215  between the first inflow port  211  and the liquid storage chamber  213 , the valve  216  between the liquid storage chamber  213  and the first outflow port  212 , the valve  217  between the liquid storage chamber  213  and the ventilation port  214 , the valve  225  between the second inflow port  221  and the curved flow path  223 , and the valve  226  between the curved flow path  223  and the second outflow port  222  will be referred to as a “valve  240 .” In the present embodiment, the valves  215 ,  216 ,  217 ,  225  and  226  have the same configuration. 
       FIG. 6A  is a schematic plan view showing a structure of the valve  240 , and  FIG. 6B  is a sectional view of the valve  240  taken along line A-A in  FIG. 6A . 
     As shown in  FIGS. 6A and 6B , the valve  240  includes a main body  241  having a pair of flow paths  243  and  244  formed therein, and a sheet-like valve body  242  fixed to an upper surface of the main body  241 . The valve body  242  is made of an elastomer and is bonded to the upper surface of the main body  241  at the peripheral portion  242   a  thereof. One end portion  243   a  of one flow path  243  and one end portion  244   a  of the other flow path  244  are opened toward the upper surface of the main body  241  at a region inward of the peripheral portion  242   a  of the valve body  242 . Normally, a lower surface of the valve body  242  is brought into close contact with the upper surface of the main body  241  by virtue of the restoration force (elastic force) of the sheet-like valve body  242 . Therefore, one end portion  243   a  of one flow path  243  and one end portion  244   a  of the other flow path  244  are closed by the valve body  242 . As a result, one flow path  243  is blocked from the other flow path  244 . 
     The operation of the valve  240  will be described with reference to  FIGS. 7A to 7C . 
     When the valve  240  is opened, first, as shown in  FIG. 7A , a tubular valve driving mechanism  249  is pressed against the upper surface of the peripheral portion  242   a  of the valve body  242 . Subsequently, as shown in  FIG. 7B , the inside of the valve driving mechanism  249  is evacuated and the region inward of the peripheral portion  242   a  of the valve body  242  is deformed so as to bulge upward. Thus, the valve body  242  is moved away from the one end portion  243   a  of one flow path  243  and one end portion  244   a  of the other flow path  244 . In the region inward of the peripheral portion  242   a  of the valve body  242 , one flow path  243  communicates with the other flow path  244 . 
     Subsequently, when the valve  240  is closed, as shown in  FIG. 7C , the inside of the valve driving mechanism  249  returns to atmospheric pressure, and the lower surface of the valve body  242  is brought into close contact with the upper surface of the main body  241  by the restoring force of the valve body  242 . Thus, one end portion  243   a  of one flow path  243  and one end portion  244   a  of the other flow path  244  are respectively closed by the valve body  242 . As a result, one flow path  243  is blocked from the other flow path  244 . 
     The cartridge for culture medium replacement use  200  configured as above can be manufactured by injection-molding a hard resin such as, for example, polystyrene or the like. In this case, the portion of the valve  240  can be formed of a hard resin and an elastomer by a two-color molding method. The method of manufacturing the cartridge for culture medium replacement use  200  is not limited to injection molding and may be manufactured by, for example, a layered modeling method using a 3D printer. 
     &lt;Culture Medium Replacement Method&gt; 
     Next, a culture medium replacement method using the cartridge for culture medium replacement use  200  will be described with reference to  FIG. 4  and  FIGS. 8A to 11D . Thick lines in  FIGS. 8A to 11D  indicate flow paths through which a liquid is passing. First, as shown in  FIG. 4 , the cartridge conveying part  109  takes out the cartridge for culture medium replacement use  200  from the cartridge storage part  107  and moves the same to the culture medium supply part  102 . 
     In the culture medium supply part  102 , as shown in  FIG. 8A , a culture medium storage container  102   a  is connected to the first inflow port  211  of the cartridge for culture medium replacement use  200 . Then, as shown in  FIG. 8B , the valve  215  between the first inflow port  211  and the liquid storage chamber  213  and the valve  217  between the liquid storage chamber  213  and the ventilation port  214  are opened. On the other hand, the valve  216  between the liquid storage chamber  213  and the first outflow port  212  is closed. In this state, a liquid culture medium is supplied from the culture medium storage container  102   a  via the first inflow port  211 . Since the valve  216  between the liquid storage chamber  213  and the first outflow port  212  is closed, the supplied liquid culture medium is stored in the liquid storage chamber  213 . Thereafter, the valve  215  between the first inflow port  211  and the liquid storage chamber  213  and the valve  217  between the liquid storage chamber  213  and the ventilation port  214  are also closed and the liquid storage chamber  213  is sealed. The valve driving mechanism  249  is installed in the culture medium supply part  102 . The valves  215  and  217  operated here are opened and closed by the valve driving mechanism  249 . 
     Subsequently, as shown in  FIG. 4 , by the cartridge conveying part  109 , the cartridge for culture medium replacement use  200  is separated from the culture medium supply part  102  in a state in which the culture medium is stored in the liquid storage chamber  213 , and is moved to the culture medium replacement part  103 . The culture container  75  is accommodated in advance in the incubator part  101  adjacent to the culture medium replacement part  103 . 
     In the culture medium replacement part  103 , as shown in  FIG. 9A , a depressurization pump is connected to the second outflow port  222  of the cartridge for culture medium replacement use  200 . In a state in which the culture container  75  is accommodated in the incubator part  101 , as shown in  FIG. 9B , the first outflow port  212  and the second inflow port  221  of the cartridge for culture medium replacement use  200  are respectively connected to the inflow port and the outflow port of the culture container  75 . Subsequently, the valve  216  between the liquid storage chamber  213  and the first outflow port  212 , the valve  217  between the liquid storage chamber  213  and the ventilation port  214 , the valve  225  between the second inflow port  221  and the curved flow path  223 , and the valve  226  between the curved flow path  223  and the second outflow port  222  are respectively opened. The curved flow path  223  is depressurized by the depressurization pump via the second outflow port  222 . As a result, the old culture medium in the culture container  75  flows out from the second inflow port  221  into the curved flow path  223 , and the new culture medium in the liquid storage chamber  213  flows into the culture container  75  from the first outflow port  212 . Thereafter, the valve  225  between the second outflow port  222  and the curved flow path  223  and the valve  226  between the curved flow path  223  and the second outflow port  222  are respectively closed, and the curved flow path  223  is hermetically sealed. The valve driving mechanism  249  is installed in the culture medium replacement part  103 . The valves  216 ,  217 ,  225  and  226  operated here are opened and closed by the valve driving mechanism  249 . 
     In the present embodiment, the culture medium replacement operation is performed while the culture container  75  is accommodated in the incubator part  101  without being taken out from the incubator part  101 . Therefore, it is possible to remarkably reduce the environmental variation with respect to the cells existing in the culture container  75 . 
     Subsequently, as shown in  FIG. 4 , by means of the cartridge conveying part  109 , the cartridge for culture medium replacement use  200  is separated from the culture medium replacement part  103  and the culture container  75  in a state in which the culture medium is accommodated in the curved flow path  223 , and is moved to the culture medium collection part  104 . 
     In the culture medium collection part  104 , as shown in  FIG. 10A , a pressurization pump (not shown) is connected to the second outflow port  222  of the cartridge for culture medium replacement use  200 . In order to supply a new culture medium to the culture container  75  and completely replace the interior of the culture container  75  with the new culture medium, it is preferable that the volume of the culture medium supplied to the culture container  75  is larger than the volume of the culture container  75 . In this regard, when the volume of the new culture medium to be supplied to the culture container  75  is larger than the volume of the culture container  75 , the new culture medium supplied from the liquid storage chamber  213  to the culture container  75  is accommodated at the side of the second inflow port  221  of the curved flow path  223 . In addition, the new culture medium and the old culture medium may be partially mixed with each other in the boundary between the new culture medium and the old culture medium. Therefore, the valve  226  between the second outflow port  222  and the curved flow path  223  and the valve  225  between the curved flow path  223  and the second inflow port  221  are respectively opened. The curved flow path  223  is pressurized from the side of the second outflow port  222  by the pressurization pump. The new culture medium accommodated at the side of the second inflow port  221  of the curved flow path  223  and the mixture of the new culture medium and the old culture medium are pushed out and discharged from the second inflow port  221  to the outside. After the entire culture medium containing the new culture medium is discharged from the second inflow port  221 , the pressurization performed by the pressurization pump is stopped. 
     Subsequently, as shown in  FIG. 10B , the culture medium analysis part  106  is connected to the second inflow port  221  of the cartridge for culture medium replacement use  200 . Thereafter, in a state in which the valve  226  between the second outflow port  222  and the curved flow path  223  and the valve  225  between the curved flow path  223  and the second inflow port  221  are respectively opened, the curved flow path  223  is again pressurized again from the side of the second outflow port  222  by the pressurization pump. As a result, the old culture medium in the curved flow path  223  is pushed out from the second inflow port  221  toward the culture medium analysis part  106  and is collected. The culture medium analysis part  106  analyzes the components of the old culture medium thus collected. The valve driving mechanism  249  is installed in the culture medium collection part  104 . The valves  225  and  226  operated here are opened and closed by the valve driving mechanism  249 . 
     Subsequently, as shown in  FIG. 4 , by means of the cartridge conveying part  109 , the cartridge for culture medium replacement use  200  is separated from the culture medium collection part  104  and the culture medium analysis part  106  and is moved to the flow path cleaning part  105 . 
     In the flow path cleaning part  105 , as shown in  FIG. 11A , a cleaning liquid storage container  105   a  is coupled to the first inflow port  211  and the second outflow port  222  of the cartridge for culture medium replacement use  200  via a three-way valve  105   b . Subsequently, as shown in  FIG. 11B , the valve  215  between the first inflow port  211  and the liquid storage chamber  213  and the valve  217  between the liquid storage chamber  213  and the ventilation port  214  are respectively opened. On the other hand, the valve  216  between the liquid storage chamber  213  and the first outflow port  212  is closed. In this state, the cleaning liquid storage container  105   a  and the first inflow port  211  are brought into communication with each other by the three-way valve  105   b  so that a cleaning liquid is supplied from the cleaning liquid storage container  105   a  via the first inflow port  211 . Since the valve  216  between the liquid storage chamber  213  and the first outflow port  212  is closed, the supplied cleaning liquid is retained inside the liquid storage chamber  213 . After the liquid storage chamber  213  is filled with the cleaning liquid, the flow path between the cleaning liquid storage container  105   a  and the first inflow port  211  is closed by the three-way valve  105   b.    
     Subsequently, as shown in  FIG. 11C , a waste liquid container  105   c  is connected to the first inflow port  211  and the second outflow port  222  of the cartridge for culture medium replacement use  200 . A pressurization pump (not shown) is connected to the ventilation port  214 . Thereafter, the valve  216  between the liquid storage chamber  213  and the first outflow port  212  is opened. The interior of the liquid storage chamber  213  is pressurized by the pressurization pump so that the cleaning liquid inside the liquid storage chamber  213  is pushed out and discharged from the first outflow port  212  to the waste liquid container  105   c.    
     Subsequently, as shown in  FIG. 11D , the valve  226  between the second outflow port  222  and the curved flow path  223  and the valve  225  between the curved flow path  223  and the second inflow port  221  are respectively opened. In this state, the cleaning liquid storage container  105   a  and the second outflow port  222  are brought into communication with each other by the three-way valve  105   b  so that the cleaning liquid is supplied from the cleaning liquid storage container  105   a  via the second outflow port  222 . The cleaning liquid supplied from the second outflow port  222  passes through the curved flow path  223  and is discharged from the second inflow port  221  to the waste liquid container  105   c . The valve driving mechanism  249  is installed in the flow path cleaning part  105 . The valves  215 ,  216 ,  217 ,  225  and  226  operated here are opened and closed by the valve driving mechanism  249 . 
     In this example, the liquid collection flow path  202  is cleaned after the liquid supply flow path  201  is cleaned. However, the liquid supply flow path  201  may be cleaned after the liquid collection flow path  202  is cleaned. 
     Subsequently, as shown in  FIG. 4 , by means of the cartridge conveying part  109 , the cartridge for culture medium replacement use  200  thus cleaned is separated from the flow path cleaning part  105  and is returned to the culture medium supply part  102 . In this way, the cartridge for culture medium replacement use  200  is reused for a subsequent culture medium replacement. 
     If the cartridge for culture medium replacement use  200  is repeatedly reused, proteins gradually adhere to and contaminate the liquid supply flow path  201  and the liquid collection flow path  202 . Therefore, by means of the cartridge conveying part  109 , the used cartridge for culture medium replacement use  200  is periodically discharged to the cartridge collection part  108 , and the unused cartridge for culture medium replacement use  200  is periodically taken out from the cartridge storage part  107 . 
     &lt;Effect&gt; 
     Next, descriptions will be made on the effects which are achieved by the present embodiment having the above-described configuration and which have not yet been described, or the effects which are particularly important. 
     According to the present embodiment, the cartridge for culture medium replacement use  200  capable of accommodating a predetermined amount of liquid culture medium moves between the culture medium supply part  102 , the culture medium replacement part  103  and the culture medium collection part  104 , whereby the culture medium replacement can be performed in a state in which the culture container  75  is separated from the culture medium supply part  102  and the culture medium collection part  104 . This eliminates the need to centrally arrange other devices such as the medium storage container  102   a  and the collection container around the culture container  75 . Thus, the liquid flow paths are not made redundant due to the physical limitation caused by device concentration. Accordingly, as compared with the conventional culture medium replacement mechanism, it is possible to shorten the liquid flow path. 
     Furthermore, according to the present embodiment, the liquid flow paths are formed in the cartridge for culture medium replacement use  200 , namely a cartridge-like part. Thus, the cartridge for culture medium replacement use  200  can be easily gripped and carried even by a commercially available robot hand. This makes it easy to automate the replacement of the liquid flow paths. 
     Moreover, according to the present embodiment, by using a plurality of cartridges for culture medium replacement use  200 , it is possible to simultaneously perform respective processes such as a culture medium supply, culture medium replacement, culture medium collection, and flow path cleaning in parallel. Therefore, it is possible to restrain the working time from being prolonged in the case of processing a plurality of culture containers  75 . 
     Furthermore, according to the present embodiment, by performing the culture medium replacement while the culture container  75  is accommodated in the incubator part  101 , it is possible to remarkably reduce the environmental variation with respect to the cells existing in the culture container  75 . In the present embodiment, it is not necessarily essential that the culture medium replacement is performed while the culture container  75  is accommodated in the incubator part  101 . The culture container  75  may be taken out from the incubator part  101  and the culture medium replacement may be performed on a culture medium replacement stage (not shown). 
     In addition, according to the present embodiment, the liquid collection flow path  202  of the cartridge for culture medium replacement use  200  includes the curved flow path  223  having a small flow path diameter and an increased flow path length. It is therefore possible to collect the old culture medium flowing out from the culture container  75  so as not to be mixed with the new culture medium along the curved flow path  223 . As a result, the old culture medium can be discriminated from the new culture medium and can be discharged from the curved flow path  223 . The components of the old culture medium old medium used inside the culture container  75  can be accurately analyzed. 
     &lt;Modification&gt; 
     Various modifications can be made with respect to the above-described embodiment. Hereinafter, modifications will be described with reference to the drawings. In the following descriptions and the drawings used in the following descriptions, the same reference numerals as those used for the corresponding parts in the above-described embodiment are used for the parts that can be configured similarly to the above-described embodiment. Duplicate descriptions will be omitted. In addition, when it is obvious that the operations and effects obtained in the above-described embodiment can be obtained in the modification, the descriptions thereof may be omitted. 
       FIG. 12A  is an internal top view showing a configuration of a first modification of the transport container capable of accommodating a plurality of culture containers  75 , and  FIG. 12B  is an internal side view of the transport container shown in  FIG. 12A . 
     The transport container  170  shown in  FIG. 12A  and  FIG. 12B  includes a housing  171  and a cassette  180  capable of holding a plurality of culture containers  75 . Among these, the housing  171  has a rectangular tubular shape with its lower end portion opened. On the other hand, the cassette  180  includes a lid body  172  fitted to the opening of the lower end portion of the housing  171  to hermetically seal the housing  171 , a support column  174  installed to extend vertically upward from the lid body  172 , and a plurality of (eight, in the illustrated example) shelves  173  installed side by side along the support column  174 . 
     Each of the shelves  173  has a disc shape and is fixed to the support column  174  in a horizontally oriented posture. On each of the shelves  173 , a plurality of (four, in the illustrated example) culture containers  75  are mounted in a rotational symmetry relationship (quadruple symmetry relationship, in the illustrated example) around the support column  174 . 
     Similar to the transport container  70  shown in  FIG. 3 , the transport container  170  shown in  FIG. 12A  and  FIG. 12B  is transported along the rail  65  to the loading part  21  of the iPS cell automatic culture devices  20 , the loading part  31  of the differentiated cell automatic culture devices  30 , or the loading part  12   a  of the iPS cell establishing device  11 , in a state in which the transport container  170  is suspended downward by the holding part  61 . 
     When taking out the culture container  75  from this transport container  170 , as shown in  FIG. 13 , the cassette  180  is pulled downward by a rotary elevator mechanism (not shown) and is taken outside of the housing  171  in a state in which the plurality of culture containers  75  is held in the cassette  180 . Then, one culture container  75  is gripped and taken out from the upper side of one shelf  173  by a robot hand (not shown). After the culture containers  75  are taken out one by one from the upper side of each of the shelves  173 , the cassette  180  is rotated by 90 degrees around the support column  174  by the rotary elevator mechanism. Then, the subsequent culture container  75  is gripped and taken out from the upper side of one shelf  173  by the robot hand. By repeating the taking-out of the culture container  75  using the robot hand and the rotation of the cassette  180  using the rotary elevator mechanism, all the culture containers  75  held in the cassette  180  can be taken out by the robot hand. 
     According to the transport container  170  configured as above, the plurality of culture containers  75  can be mounted on each of the plurality of shelves  173 . It is therefore possible to increase the number of culture containers  75  accommodated in the transport container  170  as compared with the transport container  70  shown in  FIG. 3 . This makes it possible to collectively transport a larger number of culture containers  75  into the device. 
     Furthermore, the plurality of culture containers  75  mounted on each of the shelves  173  is mounted in a rotational symmetry relationship around the support column  174 . Therefore, by rotating the cassette  180  about the support column  174 , access points of the culture containers  175  accessed by the robot hand can be made common to one point. This makes it possible to simplify the container taking-out mechanism. 
     In the example shown in  FIGS. 12A and 12B , four culture containers  75  are mounted on each shelf  173 . However, the present disclosure is not limited thereto. For example, as shown in  FIG. 14 , two culture containers  75  may be mounted on each shelf  173  in a double symmetry relationship about the support column  174 . Alternatively, n (n=3, 5, 6, 7 . . . ) culture containers  75  may be mounted on each shelf  173  in an n symmetry relationship around the support column  174 . 
     Next, a modification of the cell culture device used together with the transport container  170  shown in  FIGS. 12A and 12B  will be described. 
       FIG. 15A  is a schematic plan view showing a configuration of a modification of the cell culture device.  FIG. 15B  is a sectional view of the cell culture device taken along line B-B in  FIG. 15A .  FIG. 15C  is a sectional view of the cell culture device taken along line C-C in  FIG. 15A .  FIG. 15D  is a left side view of the cell culture device shown in  FIG. 15A . 
     The cell culture device  100   b  shown in  FIGS. 15A to 15D  includes an in-device transfer part  111  configured to take out a cassette  180  (not shown in  FIGS. 15A to 15D ) holding a plurality of culture containers  75  from a transport container  170  shown in  FIGS. 12A and 12B  and configured to transfer the cassette  180  holding the plurality of culture containers  75  up to the incubator part  101 , and a plurality of (sixteen, in the illustrated example) incubator parts  101  capable of accommodating the plurality of culture containers  75  while being held by the cassette  180 . Similar to the cell culture device  100  shown in  FIG. 4 , the cell culture device  100   b  further includes a cartridge conveying part  109  configured to convey the cartridge for culture medium replacement use  200 , a culture medium supply part  102 , a culture medium replacement part (not shown in  FIGS. 15A to 15D ), a culture medium collection part  104 , and a flow path cleaning part  105 . In  FIGS. 15A to 15D , only the portions of the first outflow port and the second inflow port of the cartridge for culture medium replacement use  200  connectable to the culture container  75  are shown. 
     As shown in  FIG. 15C , the sixteen incubator parts  101  are arranged in a 4×4 matrix shape along vertical and horizontal directions. Each of the incubator parts  101  is accessible from both the in-device transfer part  111  and the cartridge conveying part  109 . A turntable (not shown) is installed in each of the incubator parts  101  to rotate the cassette  180  holding the plurality of culture containers  75  around the support column  174 . 
     In the example shown in  FIG. 15C , a cell inspection removal part  112  is connected to the incubator part  101  located at the lower right side. The cell inspection removal part  112  takes out the culture containers  75  one by one from the incubator part  101  connected thereto. The cell inspection removal part  112  is configured to inspect the cells existing in the culture container  75  thus taken-out and remove the cells having a bad state. 
     Next, a method of using the cell culture device  100   b  shown in  FIGS. 15A to 15D  will be described. 
     First, the transport container  170  transferred by the container transfer part  60  is connected to the in-device transfer part  111 . The cassette  180  holding the plurality of culture containers  75  is taken out from the transport container  170  to the in-device transfer part  111 . 
     Subsequently, the in-device transfer part  111  loads the cassette  180  holding the plurality of culture containers  75  into one incubator part  101 . Inside the incubator part  101 , one or all of a temperature, a humidity and a gas concentration are automatically adjusted. From the viewpoint of running cost, it is preferable that the operations of other incubator parts  101  not in use are stopped at this time point. 
     Subsequently, a culture medium replacement using the cartridge for culture medium replacement use  200  is performed with respect to the culture container  75  accommodated in the incubator part  101  in the same manner as the above-described culture medium replacement method. 
     That is to say, after the cartridge for culture medium replacement use  200  conveyed by the cartridge conveying part  109  is connected to the culture medium supply part  102  and the liquid culture medium is supplied to the liquid supply flow path  201  of the cartridge for culture medium replacement use  200 , the cartridge for culture medium replacement use  200  is separated from the culture medium supply part  102 . 
     Subsequently, the cartridge for culture medium replacement use  200  conveyed by the cartridge conveying part  109  is connected to one culture container  75  mounted on one shelf  173  accommodated in the incubator part  101 . The liquid culture medium inside the liquid supply flow path  201  is supplied into the culture container  75 , and the liquid culture medium inside the culture container  75  is collected into the liquid collection flow path  202 . Thereafter, the cartridge for culture medium replacement use  200  is separated from the culture container  75 . 
     Subsequently, the cartridge for culture medium replacement use  200  conveyed by the cartridge conveying part  109  is connected to the culture medium collection part  104 . After the liquid culture medium is collected from the liquid collection flow path  202  to the culture medium collection part  104 , the cartridge for culture medium replacement use  200  is separated from the culture medium collection part  104 . 
     Subsequently, the cartridge for culture medium replacement use  200  conveyed by the cartridge conveying part  109  is connected to the flow path cleaning part  105 . The liquid supply flow path  201  and the liquid collection flow path  202  are respectively cleaned. The cartridge for culture medium replacement use  200  thus cleaned is separated from the flow path cleaning part  105  and is returned to the culture medium supply part  102 . The cleaned cartridge for culture medium replacement use  200  is used for the subsequent culture medium replacement. 
     After the culture medium replacement is performed with respect to one culture container  75  mounted on each shelf  173 , the cassette  180  is rotated by 90 degrees around the support column  174  by the turntable installed in the incubator part  101 . Then, the culture medium replacement is performed with respect to a subsequent culture container  75  mounted on each shelf  173 . 
     As described above, the plurality of culture containers  75  are accommodated in the incubator part  101  in a state in which the culture containers  75  are held by the cassette  180  and the cassette  180  is rotated around the support column  174 . Thus, in each shelf  173 , the access points of the cartridge for culture medium replacement use  200  accessed by the cartridge conveying part  109  can be made common to one point. This makes it possible to simplify the cartridge conveying part  109 . 
     The in-device transfer part  111  periodically transfers the cassette  180  holding the plurality of culture containers  75  to another incubator part  101  to which the cell inspection removal part  112  is connected. The cell inspection removal part  112  takes out the culture containers  75  one by one from the incubator part  101 . The cell inspection removal part  112  inspects the cells existing in the culture containers  75  and removes the cells having a bad state. Then, the inspected culture containers are returned into the incubator part  101 . Thereafter, the in-device transfer part  111  returns the cassette  180  holding the plurality of inspected culture containers  75  to the original incubator part  101 . 
     Meanwhile, in the cell culturing step, it is necessary to handle the plurality of culture containers  75  in order to distribute the cells into the plurality of culture containers  75  along with the growth of the cells and to continuously culture the cells. In the case where the culture containers  75  are transferred one by one inside the device as in the conventional cell culture device, even if the cells have the same basis, a time difference occurs between the initial culture container  75  and the final culture container  75  as the culture proceeds. This greatly changes the culture conditions. 
     On the other hand, according to the cell culture device  100   b  shown in  FIGS. 15A to 15D , the plurality of culture containers  75  is collectively conveyed in a state in which the culture containers  75  are held by the cassette  180 . It is therefore possible to reduce a difference in culture time. 
     In a case where the subculture of cells are performed in the cell culture device  100   b  shown in  FIGS. 15A to 15D , the number of incubator parts  101  to be used may be increased in conformity with the increasing number of culture vessels. By gradually increasing the number of incubator parts  101  to be used, it is possible to reduce the running cost. 
     Finally, the foregoing descriptions of the embodiment and the disclosure of the drawings are nothing more than one example for describing the present disclosure recited in the claims. The present disclosure recited in the claims shall not be limited by the foregoing descriptions of the embodiment and the disclosure of the drawings. In addition, the respective embodiments can be appropriately combined unless the processing contents are inconsistent. 
     
       
         
           
               
             
               
                   
               
               
                 EXPLANATION OF REFERENCE NUMERALS 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 75: culture container, 
                 100, 100b: cell culture device 
               
               
                 101: incubator part 
                 102: culture medium supply part 
               
               
                 103: culture medium replacement part 
                 104: culture medium collection part 
               
               
                 105: flow path cleaning part 
                 106: culture medium analysis part 
               
               
                 107: cartridge storage part 
                 108: cartridge collection part 
               
               
                 109: cartridge conveying part 
                 200: cartridge for culture medium replacement use 
               
               
                 201: liquid supply flow path 
                 202: liquid collection flow path 
               
               
                 211: first inflow port 
                 212: first outflow port 
               
               
                 213: liquid storage chamber 
                 214: ventilation port 
               
               
                 221: second inflow port 
                 222: second outflow port 
               
               
                 223: curved flow path 
                 224: tank-shaped flow path