Abstract:
A cell culture apparatus is provided in which a plurality of culture vessels can be mounted and removed in a set, which has a regulation mechanism that can fix each culture vessel in the correct position, and which enables suppression of production costs. The cell culture apparatus has one or a plurality of concave portions, and can arrange and accommodate one or a plurality of culture vessels in the concave portions. Spring-type fixing devices are provided in the depth direction and the width direction, respectively, in each concave portion. The two spring-type fixing devices press against side surfaces of the culture vessel to fix the culture vessel in the culture vessel set. At this time, by pressing against the culture vessel at two points, the position of the culture vessel at the time of fixing can be maintained with good accuracy.

Description:
CLAIM OF PRIORITY 
       [0001]    The present application claims priority from Japanese application JP 2006-338067 filed on Dec. 15, 2006, the content of which is hereby incorporated by reference into his application. 
       CROSS REFERENCE TO RELATED APPLICATIONS 
       [0002]    U.S. patent application Ser. Nos. 10/767,993, 11/192,023, 11/774,093 and 11/756,015 are co-pending applications of the present application, the contents of which are incorporated herein by cross-reference. 
       BACKGROUND OF THE INVENTION 
       [0003]    1. Field of the Invention 
         [0004]    The present invention relates to a cell culture apparatus that cultures cells using a culture vessel, and a method that controls the cell culture apparatus. 
         [0005]    2. Background Art 
         [0006]    Conventionally, cell culture work is performed manually by skillful workers in a clean room that has been completely sterilized. Therefore, when culturing cells in large amounts in preparation for industrialization, time and expense is required to educate and train workers and the workload of the workers increases, and there is a possibility of human errors or taking of samples mistakenly and also of contamination caused by humans who possess fungi or the like. A large amount of costs are incurred to provide countermeasures for these possibilities. This represents a large barrier with respect to industrialization. These problems can be solved by automating a series of culture operations. With respect to automation of a step in cell culturing, JP Patent Publication (Kokai) No. 2006-149237 describes a case of automating a step in which a culture medium is introduced into a culture vessel such as for seeding of cells or exchanging the medium. In the method of connecting a joint and a culture vessel described in JP Patent Publication (Kokai) No. 2006-149237, a robot manipulator is used for attaching and detaching the culture vessel and the joint on the manipulator side. The configuration is such that, at that time, a connection can be made in a state in which a clean state is maintained in which a leakage of culture medium is prevented between the inside of the culture vessel and a tube of the joint through a valve by a resin membrane. Further exchange of the medium is carried out by supplying the culture medium from the bottom and discharging the medium from the top in a state in which the culture vessel that is connected by the manipulator is stood in the vertical direction. 
         [0007]    Further, a method may be considered which makes it possible to rapidly and efficiently transfer a culture medium from a plurality of culture vessels, reduce loss of the culture medium and readily carry out washing. One example thereof is described in JP Patent Publication (Kokai) No. 2004-89126. In this case, a culture vessel is provided in which a plurality of culturing holes for introducing a culture medium are formed. Through holes that penetrate through the culture vessel and open on the outer surface side of the culture vessel are formed in the bottom of these culturing holes, and means is provided that forcibly discharges the culture medium in the culturing holes from the through holes. 
       SUMMARY OF THE INVENTION 
       [0008]    Cell transplantation may involve autotransplantation in which cells that were extracted from an individual are treated in vitro and returned to the same individual or allotransplantation in which cells that were extracted from another individual such as in the case of a body donation are treated in vitro and transplanted to an individual of the same type. For autotransplantation, because it is only necessary to produce a quantity of tissue that is sufficient for treating the affected area of the relevant individual and because the amount of cells that can be extracted is also small, culturing is performed using one or a plurality of culture vessels. In contrast, for allotransplantation, since many cells that are supplied by a body donation or the like are used and that number of cells is increased in large quantities to produce a large amount of tissue, culturing is performed using many culture vessels. 
         [0009]    However, when culture vessels are treated one at a time in a step of exchanging medium or seeding of cells in automated culturing for allotransplantation or the like, a large amount of time is required. This becomes a problem not only with respect to costs, but also with respect to quality control since the activity of the cultures or cells is not uniform and other environmental stresses also act on the cells. Therefore, to reduce the treatment time for seeding of cells or exchanging medium, a method is required that treats a plurality of culture vessels at the same time. Further, in this case it is also necessary that there are no variations with respect to the positioning of each of the plurality of culture vessels. Furthermore, when accommodating a plurality of culture vessels, since the weight increases in comparison to the case of a single culture vessel, the power of the motor of a conventional manipulator is too small. Hence, it is necessary to exchange the motor for a more powerful one, and it is not possible to suppress the cost of the cell culture apparatus by making use of a manipulator that is suited for a single culture vessel. 
         [0010]    The present invention was made in consideration of the above circumstances, and an object of the present invention is to provide a cell culture apparatus in which a plurality of culture vessels can be mounted and taken off in a set and also has a regulation mechanism that enables each culture vessel to be fixed in the correct position. A further object of the present invention is to provide a cell culture apparatus that suppressed costs. 
         [0011]    To solve the above described problems, according to the present invention there is provided a culture vessel set having one or a plurality of concave portions in which one or a plurality of culture vessels can be arranged and accommodated. In the concave portions of the culture vessel set, a spring-type fixing device is provided in the depth direction and the width direction, respectively. The two spring-type fixing devices press against the sides of the culture vessel to fix the culture vessel set. At this time, by being pressed at two points, it is possible to retain the position of the culture vessel with good accuracy at a time of fixing. A hole with a valve is provided on the top surface of the culture vessel for connecting to the culture space inside the culture vessel, and when a culture vessel is fixed in the culture vessel set, a culture medium can be supplied from the top surface thereof. 
         [0012]    A cell culture apparatus is provided that comprises a manipulator having a joint that is connected with flow channels to simultaneously supply a culture medium to inside a culture vessel set in which one or a plurality of culture vessels are fixed. Tubes that are connected to the flow channels extrude from the lower portion of the joint in a joint portion of the manipulator, and those tubes connect to holes of the culture vessels to enable supply of a liquid to inside the culture vessels that are held in a vertical direction by the manipulator. The culture vessel set is sandwiched between a rack and the joint at that time, and by rotating the rack the entire structure comprising the culture vessel set, the joint, and the rack can be stood up vertically. By placing the entire structure in the horizontal direction after supplying the liquid and then detaching the joint, seeding of cells or exchange of a medium can be realized. 
         [0013]    More specifically, a cell culture apparatus according to the present invention is an apparatus that cultures cells using culture vessels, comprising a culture vessel set having a plurality of concave portions for accommodating a plurality of culture vessels, wherein each of the plurality of concave portions have pressurization means that is disposed in at least one portion of an inner wall and that pressurizes the culture vessels that is accommodated. 
         [0014]    Further, the cell culture apparatus according to the present invention is a cell culture apparatus that cultures cells using culture vessels, comprising: a culture vessel set that holds a plurality of culture vessels; a mounting rack for mounting the culture vessel set; a joint having at least one liquid supply means that supplies a liquid to the plurality of culture vessels and at least one liquid recovery means that discharges a liquid from the plurality of culture vessels; first coupling means that couples a liquid inlet portion of the plurality of culture vessels and the liquid supply means; second coupling means that couples a liquid outlet portion of the plurality of culture vessels and the liquid recovery means; and a manipulator for connecting the liquid supply means and the liquid recovery means to the plurality of culture vessels through the first and the second coupling means, that moves the joint and inserts the culture vessel set between the joint and the mounting rack. The cell culture apparatus further comprises state changing means that changes the culture vessel set, the joint, and the mounting rack that are integrated into one piece from a horizontal state to a vertical state, wherein the liquid supply means and the liquid recovery means operate in a vertical state. Further, the state changing means is provided in the mounting rack and comprises a first rotating shaft and a drive portion for rotating the culture vessel set, the joint, and the mounting rack that are integrated into one piece, the joint has a second rotating shaft that rotates in an integrated condition with the first rotating shaft, and the mounting rack has a bearing portion that accommodates the second rotating shaft in an integrated condition with the first rotating shaft. The first rotating shaft is provided on a center line of the mounting rack and the second rotating shaft is provided on a center line of the joint. 
         [0015]    The present invention also provides a control method of a cell culture apparatus. In this case the cell culture apparatus comprises: a culture vessel set that holds a plurality of culture vessels; a mounting rack for mounting the culture vessel set; a joint having at least one liquid supply means that supplies a liquid to the plurality of culture vessels and at least one liquid recovery means that discharges a liquid from the plurality of culture vessels; first coupling means that couples a liquid inlet portion of the plurality of culture vessels and the liquid supply means; second coupling means that couples a liquid outlet portion of the plurality of culture vessels and the liquid recovery means; a manipulator for moving the joint, first detection means that detects that the culture vessel set is mounted in the mounting rack; and control means that controls an operation of the cell culture apparatus. 
         [0016]    For this cell culture apparatus, the control method comprises a first step in which the control means moves the manipulator based on a detection result obtained by the detection means, inserts the culture vessel set between the joint and the mounting rack, and connects the liquid supply means and the liquid recovery means to the plurality of culture vessels through the first and the second coupling means. The cell culture apparatus further comprises second detection means that detects that the culture vessel set is inserted in an integrated condition between the joint and the mounting rack, and state changing means that changes an orientation of the culture vessel set, the joint, and the mounting rack that are integrated into one piece. For this cell culture apparatus, the control method further comprises a second step in which the control means controls the state changing means so as to place the culture vessel set, the joint, and the mounting rack that are integrated into one piece into a vertical state from a horizontal state based on a detection result of the second detection means, and a third step in which the control means causes the liquid supply means and the liquid recovery means to operate in a vertical state. 
         [0017]    The state changing means is provided in the mounting rack and comprises a first rotating shaft and a drive portion for rotating the culture vessel set, the joint, and the mounting rack that are integrated into one piece. The joint has a second rotating shaft that rotates in an integrated condition with the first rotating shaft. The mounting rack has a bearing portion that accommodates the second rotating shaft in an integrated condition with the first rotating shaft. The detection means is configured to detect that the second rotating shaft is accommodated in the bearing portion. 
         [0018]    Further features of the present invention will be apparent from the description of the preferred embodiments for implementing the present invention and the attached drawings as described hereunder. 
         [0019]    According to the cell culture apparatus of the present invention, a plurality of culture vessels can be mounted and dismounted in a set, and each culture vessel can be fixed in the correct position. Further, the cost of the cell culture apparatus can be suppressed by making use of a conventional manipulator. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0020]      FIG. 1  is a view that illustrates an embodiment of the present invention, and shows an overall view relating to connection of a culture vessel and a culture vessel set; 
           [0021]      FIG. 2  is a view that illustrates an embodiment of the present invention, and shows a configuration diagram of the top surface when connecting to a culture vessel set that has one or a plurality of concave portions inside and which can fix a culture vessel by means of two spring-type fittings; 
           [0022]      FIG. 3A  is a view that illustrates the configuration of a spring-type fixing device for fixing a culture vessel; 
           [0023]      FIG. 3B  is a conceptual diagram that illustrates a method for fixing a culture vessel using a spring-type fixing device; 
           [0024]      FIG. 4  is a conceptual diagram that illustrates a method for taking out a culture vessel that is fixed in the culture vessel set; 
           [0025]      FIG. 5  is an overall configuration diagram of a manipulator that connects a joint that is connected to flow channels to a culture vessel; 
           [0026]      FIG. 6A  is a view showing a state in which a culture vessel set is mounted on a culture vessel set rack (mounting rack); 
           [0027]      FIG. 6B  is a view showing a state in which the culture vessel set, the culture vessel set rack, and the joint are integrated into one piece; 
           [0028]      FIG. 6C  is a view showing a state in which the culture vessel set, the culture vessel set rack, and the joint that are integrated into one piece are placed in a vertical state; 
           [0029]      FIG. 7A  is a view showing a state before the joint and the culture vessel set are connected, as viewed from the side; 
           [0030]      FIG. 7B  is a view showing a state in which the joint and the culture vessel set are connected, as viewed from the side; 
           [0031]      FIG. 8A  is a view showing a state before one culture vessel and a flow channel of the joint are connected; and 
           [0032]      FIG. 8B  is a view showing a state in which one culture vessel and a flow channel of the joint are connected. 
       
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
       [0033]    Hereunder, an embodiment of the present invention is described in detail with reference to the attached drawings. However, this embodiment represents no more than one example for implementing the present invention, and it should be understood that the embodiment does not limit the present invention. 
       &lt;Cell Culture Apparatus&gt; 
       [0034]    A feature of the cell culture apparatus that is used with the present invention is, as described later, the configuration and operation of a culture vessel  10 , a culture vessel set  20 , and a joint  50  that performs the supply and discharge of a liquid into and out of each culture vessel. The configuration of a conventional cell culture apparatus can be used for the remaining configuration, and therefore the remaining configuration is not shown. However, the minimum configuration of the cell culture apparatus will now be described as a precaution. The cell culture apparatus comprises a culture chamber; a culture vessel set introduction portion for introducing the culture vessel set  20  containing a plurality of the culture vessels  10  into a culture chamber and for removing it therefrom; conveying means that conveys the culture vessel set  20  between the culture vessel set introduction portion and a predetermined culturing position; and a manipulator  51  that handles the culture vessel set  20  at the predetermined culturing position. 
       &lt;Culture Vessel and Culture Vessel Set&gt; 
       [0035]      FIG. 1  is an overall view of a state when connecting the culture vessel  10  and the culture vessel set  20 , which best illustrates the features of the present invention.  FIG. 2  is a top view of the configuration of the culture vessel set  20 .  FIGS. 3A  and B are schematic diagrams of a spring-type fixing device  22  when placing the culture vessel  10  in the culture vessel set  20 .  FIG. 4  is a schematic diagram relating to a method of taking the culture vessel  10  out from the culture vessel set  20 . 
         [0036]    The configurations of the culture vessel  10  and the culture vessel set  20  as shown in  FIG. 1 ,  FIG. 2 ,  FIG. 3 , and  FIG. 4  and the connection thereof will now be described. 
         [0037]    First, the configurations of the culture vessel  10  and the culture vessel set  20  as shown in  FIG. 1  are described. The culture vessel  10  has a culture space  11  on the inside for culturing cells with a closed system. A culture medium is inserted therein to culture cells. A pair of connection holes  12  for supplying and discharging a culture medium from and to the outside are provided on the top surface of the culture vessel  10 . When supplying a culture medium, the culture medium is allowed to flow from one of the connection holes  12 , and the culture medium and gas is discharged from the other connection hole  12 . The culture vessel set  20  has one or a plurality of concave portions  21 , and the culture vessel  10  is inserted into one of the concave portions  21 . At that time, two spring-type fixing devices  22  that are inside the concave portion  21  are used to fix the culture vessel  10 . The spring-type fixing devices  22  press against the culture vessel side surface  13  to fix the culture vessel  10 . A dismounting hole  23  for taking out the culture vessel  10  is also provided in the culture vessel set  20 , and a fitting  41  can be inserted therein to take out the culture vessel  10  of interest. The dismounting hole  23  penetrates the bottom surface of the culture vessel set  20 . 
         [0038]    The configuration of the culture vessel set  20  will now be described using  FIG. 2 . Two spring-type fixing devices  22  and one dismounting hole  23  are provided for each concave portion  21  of the aforementioned culture vessel set  20 . The spring-type fixing devices  22  are fixed to the bottom surface or the side surface of the concave portion  21  of the culture vessel set  20 . The spring-type fixing devices  22  are each provided in the vertical direction, and a raw material having the strength of stainless steel is used so that the spring-type fixing devices  22  do not rust in a high humidity environment when culturing. When the culture vessel  10  is set in the concave portion  21 , by the spring-type fixing devices  22  pushing the culture vessel side surfaces  13  with forces  24  from two directions, the positioning accuracy in the horizontal direction of the culture vessel  10  when setting the culture vessel  10  can be enhanced. 
         [0039]    Further, a force is applied to all the culture vessel side surfaces  13  thereby, and it is therefore possible to prevent the culture vessel  10  from moving in the vertical direction when an external force is applied or from detaching from the culture vessel set  20 . By providing two of the spring-type fixing devices  22  at side surface portions that adjoin each other in each concave portion  21  as described above, each culture vessel  10  can be positioned with good accuracy at an angle portion A (see  FIG. 2 ) of the concave portion  21 . Thus, since the culture vessel  10  is pressingly supported at four surfaces by the spring-type fixing devices  22  and the side surfaces on which the spring-type fixing devices  22  are not disposed in the concave portion  21 , disposition errors (displacement) of the culture vessel  10  can be suppressed to a minimum. In this connection, the disposition positions of the two spring-type fixing devices  22  need not necessarily be the positions shown in  FIG. 2 , and any side surface may be used as long as the spring-type fixing devices  22  make an L-shape and are at the same positions in all of the concave portions  21 . 
         [0040]    The configuration of the spring-type fixing device  22  will now be described in more detail using  FIG. 3 .  FIG. 3A  shows the state before connecting the culture vessel  10 .  FIG. 3B  shows the state after connecting the culture vessel  10 . As shown in  FIG. 3A , the spring-type fixing device  22  is strongly fixed with a screw  25  or the like to the bottom surface or side surface of the concave portion  21  of the culture vessel set  20 . The screw  25  is composed of a raw material that does not rust, such as stainless steel or a resin. Further, although the spring-type fixing devices  22  are in a free state before connection of the culture vessel  10 , after connecting the culture vessel  10 , as shown in  FIG. 3B , by pressing the culture vessel side surface  13  with the force  24  with the spring-type fixing device  22 , a high positioning accuracy can be maintained in the horizontal direction as described above. Movement in the vertical direction can also be prevented by the frictional force of the pressing portion  26 . More specifically, each culture vessel  10  is firmly held in each concave portion  21  of the culture vessel set  20  by the force  24  (force in a direction that is substantially parallel to the bottom surface of the concave portion  21 ) that is generated by a repulsive force of the spring-type fixing device  22  and a frictional force (force in a direction that is substantially perpendicular to the bottom surface of the concave portion  21 ) between the spring-type fixing device  22  and the culture vessel  10  that is generated at the pressing portion  26 . 
         [0041]    An operation for removing the culture vessel  10  from the culture vessel set  20  will now be described using  FIG. 4 . First, a tip  42  of a removal fitting  41  having a convex portion in a shape that matches a removal hole  23  of the culture vessel set  20  is inserted into the removal hole  23 . Next, the inserted fitting  41  is pushed out from the bottom surface of the culture vessel set  20  towards the top surface thereof. Thus, since the culture vessel  10  is displaced from the standard position of the concave portion  21  of the culture vessel set  20 , it is easy to remove the culture vessel  10 . To enable sterilization, the removal fitting  41  is composed of a material that is water resistant, temperature resistant, and gas resistant. 
         [0042]    In this connection, the removal operation may be implemented by, for example, removing the culture vessel set  20  to outside of the cell culture apparatus from the culture vessel set introduction portion, after which the operator (for example, physician or the like) inserts the removal fitting  41  into the removal hole  23  to remove a culture vessel  10  at a stage when the respective culture vessel  10  is to be used. Alternatively, the removal operation may be implemented by adopting a configuration in which a desired culture vessel is automatically disengaged from the concave portion  21  at the culture vessel set introduction portion upon an instruction from an operator. 
       &lt;Configuration and Operation of Manipulator and Connection of Joint and Culture Vessel&gt; 
       [0043]      FIG. 5  is a configuration diagram of a manipulator  51  having a joint  50  for connecting to the culture vessel set  20  and supplying a liquid into the culture vessel  10 .  FIGS. 6A to 6C  are schematic diagrams that illustrate an operation that connects the culture vessel set  20  and the manipulator  51 .  FIGS. 7A and 7B  are views that illustrate in detail an operation that connects the joint  50  and the culture vessel set  20 .  FIGS. 8A and 8B  are schematic drawings that relate to connection of flow channels  52  of the joint  50  and the culture vessels  10 . 
         [0044]    The configuration of the manipulator  51  having the joint  50  for supplying a liquid into the culture vessel  10  of the culture vessel set  20  will now be described using  FIG. 5 . The manipulator  51  has a joint  50  and flow channels  52  that are connected thereto; a rotating shaft  53  for rotating the joint  50 ; a stopper  54  for restraining the rotating shaft  53  so that the rotating shaft  53  does not rotate; a motor  55  for causing the joint  50  to move vertically (in the direction of an arrow  63 ); a culture vessel set rack  56  for mounting the culture vessel set  20  thereon; a rotating shaft  57  for rotating the culture vessel set rack  56  and the joint  50 ; an arm (joint rotating shaft accommodating means: bearing portion)  58 ; and a connection groove  59  in which a rotating shaft  53  of the joint  50  is accommodated. The manipulator  51  inserts the culture vessel set  20  between the joint  50  and the culture vessel set rack  56 , and rotates all of these parts together in the vertical direction from the horizontal direction by rotating the rotating shaft  57  of the culture vessel set rack  56  using the motor  60 . At this time, a vertical state is detected by a sensor  61  such as an encoder, and based on the detection result an unshown control portion for controlling the entire cell culture apparatus stops the operation of the motor  60 . This mechanism makes it possible to supply a culture medium to the culture vessels  10  in a vertical state. 
         [0045]    In a state in which the rotating shaft  53  of the joint  50  is not set in the arm  58 , the stopper  54  acts to retain the joint  50  in a horizontal state. When a sensor  74  detects that the rotating shaft  53  is set in the arm  58 , the aforementioned control portion releases the stopper  54  to thereby enter a state in which the rotating shaft  53  can be rotated by the motor  60 . The rotating direction is detected by the sensor  61 . Further, when the sensor  74  detects that the rotating shaft  53  is taken out from the arm  58 , the control portion causes the stopper  54  to act again on the rotating shaft  53  to retain the joint  50  in a horizontal state so that it does not wobble. 
         [0046]    Next, connection of the culture vessel  10  and the joint  50  will be described using  FIG. 6  and  FIG. 7 . As shown in  FIG. 6A , first the culture vessel set  20  is mounted onto the culture vessel set rack  56 . According to the present embodiment, the culture vessel  10  is mounted in a state in which connection holes  12  of the culture vessels  10  face the top surface and the two connection holes provided in each culture vessel  10  are parallel with the X axis. Further, the joint  50  is disposed so that the flow channels  52  come over the connection holes  12  so that they can be connected to the connection holes  12 . 
         [0047]    A sensor  62  is provided in the culture vessel set rack  56 . The sensor  62  can detect that the culture vessel set  20  is mounted on the rack  56 . When the sensor  62  detects that the culture vessel set  20  is mounted on the culture vessel set rack  56 , the aforementioned control portion operates the motor  55  to slide the joint  50  in the vertical direction (arrow  63 ) and connect the connection holes  12  of the culture vessels  10  and the flow channels  52  of the joint  50 . This state is shown in  FIG. 6B . 
         [0048]    The details of the operation that connects the joint  50  and the culture vessel  10  will be now described further using  FIG. 7 .  FIG. 7A  shows the state before connection. As shown in  FIG. 7A , the rotating shaft  53  is provided at substantially the center of a side surface portion of the joint  50 . The rotating shaft  53  is connected to the manipulator  51  through a joint arm  71 . Because the joint arm  71  is moved upward and downward by the motor  55  of the manipulator  51 , the joint  50  can also move upward and downward (arrow  72 ). Further, the joint  50  can rotate (arrow  73 ) from the horizontal direction to the vertical direction by rotation of the rotating shaft  53 . As described above, the stopper  54  is provided so that the joint  50  does not freely rotate, and can be fixed in a predetermined position (before connection, the position is a horizontal state). 
         [0049]    The arm (joint rotating shaft accommodating portion)  58  is provided in the culture vessel set rack  56 . The rotating shaft  57  that connects with the motor  60  is provided in the arm  58 . When the rotating shaft  53  of the joint  50  is mounted in the connection groove  59  of the arm  58 , the rotating shaft  57  and the rotating shaft  53  collinearly overlap. 
         [0050]    First, when the sensor  62  detects that the culture vessel set  20  is mounted on the culture vessel set rack  56 , the aforementioned control portion operates the motor  55 . Subsequently, the joint  50  descends and the rotating shaft  53  of the joint  50  moves to the connection groove  59 . Further, the sensor  74  inside the connection groove  59  detects that the rotating shaft  53  is inside the connection groove  59 , and based on that detection result the operation of the motor  55  is stopped by the control portion. When the rotating shaft  53  of the joint  50  is accommodated in the connection groove  59  of the arm  58 , as shown in  FIG. 7B , the state is such that the respective rotating shafts  53  and  57  are disposed on the same line and the joint  50  and the culture vessels  10  are connected (state in which the connection holes  12  and the flow channels  52  fit together). Subsequently, when the rotating shaft  57  of the motor  60  that is connected to the arm  58  is rotated (arrow  73 ), the rotating shaft  53  of the joint  50  also rotates (arrow  73 ) in a similar manner, and the joint  50  and the culture vessels  10  can be rotated (arrow  73 ) in a state in which the culture vessel set  20  and the joint  50  are held in a connected state on the culture vessel set rack  56 . 
         [0051]      FIG. 6C  shows a state in which the joint  50  and the culture vessels  10  are rotated (arrow  73 ) from the horizontal direction to the vertical direction. A culture medium is fed into the flow channels and supplied to the culture vessels  10 . The configuration is such that, at this time, the culture medium is supplied from the connection holes  12  at the lower part of the culture vessel  10  and waste liquid or discharge air flows out from the connection holes  12  in the upper part of the culture vessel  10  that are paired with the connection holes  12  at the lower part thereof. After supply of the culture medium is completed, the joint  50  and the culture vessels  10  can be returned to the horizontal position to reach the state shown in  FIG. 6B , the joint  50  can be detached from the culture vessels  10 , and as shown in the completely separated state shown in  FIG. 6A , exchange of the culture medium or filling of cells in a culture medium can be ended. 
         [0052]    Next, the operations before and after connection of the culture vessels  10  and the flow channels  52  of the joint  50  as described above will be described in detail using  FIG. 8 . In  FIG. 8A , a connection between the joint  50  and the flow channels  52  is made with connectors  80 . A tube  81  protrudes from the tip of each connector  80 . At the bottom surface of the joint  50  are provided a resin membrane (for example, silicon resin)  82  and a micro-projection  83  that is made with the same material as the resin membrane  82 . Further, a resin valve  84  is affixed to the top surface of the culture vessel  10  to fulfill the role of a valve when the connection hole  12  and the tube  81  are connected. 
         [0053]    Further, as shown in  FIG. 8B , in a state in which the joint  50  and the culture vessels  10  are connected, the tube  81  is configured to push aside the resin valve  84  to connect the connection hole  12  and the flow channel  52 . At this time, since the micro-projection  83  is designed to be somewhat larger than the connection hole  12 , together with the action of the resin valve  84 , leakage can be prevented when supplying a liquid. When a liquid is to be supplied, the joint  50 , the culture vessel set  20 , and the culture vessel set rack  56  that are integrated into one piece are controlled so as to rotate (arrow  73 ) from the horizontal direction to the vertical direction. Subsequently, at the pair of connection holes  12 , a liquid  86  is supplied from below and discharge liquid and discharge air  87  are discharged from above. It is thereby possible to efficiently supply a liquid to inside of the culture vessels  10 , and situations in which air bubbles are left in the culture space of the culture vessels  10  are eliminated. 
         [0054]    After supply of the liquid is completed, the joint  50 , the culture vessel set  20 , and the culture vessel set rack  56  that are integrated into one piece are rotated from the vertical direction to the horizontal direction to be returned to a horizontal state. The joint  50  is then detached from the culture vessel set rack  56 . Subsequently, the culture vessel set  20  and the culture vessels  10  are detached from the culture vessel set rack  56  as required. 
         [0055]    It is thus possible to realize a highly flexible system that can supply a liquid for one or a plurality of culture vessels  10  or culture vessels  10  that are of different shapes or the like by simply changing the shape of the culture vessel set  20  and the joint  50 . Further, by providing two of the spring-type fixing devices  22  for each culture vessel  10 , it is possible to simply and easily enhance the positioning accuracy of the culture vessels  10  and prevent the culture vessels  10  from popping out. 
       &lt;Summary&gt; 
       [0056]    As described above, according to the embodiment of the present invention, one or a plurality of culture vessels can be fixed by a culture vessel set while maintaining high positional accuracy. Further, a conventional manipulator can be adapted to correspond to various culture conditions by merely changing the configuration (shape) of the joint and the culture vessel set. The present cell culture apparatus can also correspond to a case in which culture vessels differ for each tissue that is the object of culturing. Since it is possible to make use of the important component parts of a conventional manipulator to realize new functions in this manner, the cost of the cell culture apparatus can be suppressed to the utmost. Furthermore, with respect to automation of cell culture work in which a high degree of cleanliness by elimination of human workers is being demanded, since the cell culture apparatus according to the above described embodiment is flexible, can support a large quantity of culture vessels, and can handle the culture vessels with high positional accuracy, it is anticipated that there will be a high demand in industries dealing with areas such as cell transplants and regenerative medicine and that the utilization possibility is high. 
         [0057]    According to the cell culture apparatus of the present embodiment, a culture vessel set having a plurality of concave portions for accommodating a plurality of culture vessels is proposed. Each of the plurality of concave portions of the cell culture set has pressurization means (for example, comprising a plate spring) that is provided in at least one portion of an inner wall and that pressurizes the culture vessel that is accommodated therein. It is thereby possible to fix a plurality of culture vessels at set positions in the culture vessel set. 
         [0058]    The concave portions each form a square shape having four side surfaces, and the pressurization means is disposed on two adjoining side surfaces of each concave portion. It is therefore possible to stably retain the culture vessel by means of the two pressurization means and the two side surfaces on which the pressurization means is not disposed. As described above, the pressurization means, for example, comprises a plate spring that presses against a side surface of a culture vessel, and the culture vessel is fixed by a repulsive force of the plate spring in a substantially parallel direction (lateral direction) with respect to the bottom surface and by a frictional force of the plate spring in a substantially vertical direction (longitudinal direction) with respect to the bottom surface. It is thus possible to exert a retaining force in the direction of the XY plane and also in the Z-axis direction, so that the accuracy in all directions can be maintained. Thus, since the culture vessels can be positioned at a fixed position by the pressurization means and the side surfaces of the concave portions, it is possible to improve the handling position accuracy in the manipulator. 
         [0059]    Vessel removal holes are provided in the bottom surface of the culture vessel set. The retention force on a culture vessel can be released and the culture vessel can be removed by inserting a vessel removal member (fitting) into a vessel removal hole and pushing the vessel removal member in the upward direction. Through this removal method, a culture vessel can be easily removed even when there is a strong retention force. Further, loading a culture vessel is also simple since a spring force is utilized. It is thus possible to load the required number of culture vessels in the culture vessel set before treatment, and to remove and use only the required number of culture vessels from the culture vessel set after treatment. 
         [0060]    Further, the cell culture apparatus according to the embodiment of the present invention comprises: a culture vessel set that holds a plurality of culture vessels; a mounting rack for mounting the culture vessel set; a joint having at least one liquid supply means that supplies a liquid to the plurality of culture vessels and at least one liquid recovery means that discharges a liquid from the plurality of culture vessels; first coupling means that couples a liquid inlet portion of the plurality of culture vessels and the liquid supply means; second coupling means that couples a liquid outlet portion of the plurality of culture vessels and the liquid recovery means; and a manipulator for moving the joint and inserting the culture vessel set between the joint and the mounting rack in an integrated condition to connect the liquid supply means and the liquid recovery means to the plurality of culture vessels through the first and the second coupling means. Since the culture vessel set is inserted between the mounting rack and the joint in this manner, it is possible to prevent leaks from the culture vessels. 
         [0061]    The culture vessel set, the joint, and the mounting rack that are integrated into one piece are shifted from a horizontal state to a vertical state, and supply and recovery of a liquid with respect to the culture vessels is performed in that state. Since these operations are performed in a vertical state, air bubbles are not generated inside the culture vessels. Shifting of the culture vessel set, the joint, and the mounting rack from a horizontal state to a vertical state is performed by rotating with a motor. The motor is disposed in the mounting rack. A rotating shaft is also provided in the joint. The rotating shaft of the joint is fitted into a bearing that is provided in the mounting rack. The rotating shaft of the mounting rack and the rotating shaft of the joint are integrated into one piece and are rotated by the aforementioned motor, and as a result the culture vessel set, the joint, and the mounting rack that are integrated into one piece rotate and shift from a horizontal state to a vertical state. It is thus possible to simultaneously supply a culture medium to one or a plurality of culture vessels, and treatment can be performed efficiently. 
         [0062]    In this connection, the rotating shaft of the mounting rack is provided on the center line of the mounting rack, and the rotating shaft of the joint is also provided on the center line of the joint. Thus, in comparison to a case in which the rotating shaft is at an end of a joint (see JP Patent Publication (Kokai) No. 2006-149237), since it is easier to maintain balance when shifting the culture vessel set and the joint from a horizontal state to a vertical state and it is not necessary to boost the output of the motor, a conventional manipulator for a single culture vessel can be diverted for use as a manipulator for a plurality of culture vessels. Thereby, the cost of the cell culture apparatus can be reduced. 
         [0063]    Further, although the joint is in a horizontal state when it is not integrated as one piece with the mounting rack and the culture vessel set, a stopper that immobilizes the joint is provided to maintain that state. This stopper is not released unless the joint is integrated as one piece with the mounting rack and the culture vessel set (unless the rotating shaft of the joint is fitted into the bearing of the mounting rack). Accordingly, a connection tip portion of flow channels that form liquid supply/recovery means can be firmly connected to a liquid inlet/outlet portion of a culture vessel.