Patent Abstract:
A cell culture device comprises a fluid mechanism for branching a liquid appropriately in its flow passage when the liquid is injected to a plurality of culture vessels in a set and feeding equal amounts of the culture broth to the culture vessels at the same time. The device, in one form, has one or plural recess portions so that one or plural culture vessels can be contained in alignment, in view of space efficiency, m x n (row x column) pieces of culture vessels can be set in a matrix type. By using a pump, the culture broth is branched by a multi-way valve for distributing the liquid in an appropriate amount from a tank containing the culture broth, and the liquid is fed at the same time to the plurality of culture vessels in the culture vessel set.

Full Description:
CLAIM OF PRIORITY 
     The present application claims priority from Japanese patent application JP 2007-305237 filed on Nov. 27, 2007, the content of which is hereby incorporated by reference into this application. 
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The present invention relates to a cell culture device for culturing a cell using a culture vessel. 
     2. Background Art 
     Cell culture work has been carried out by a skilled worker in a clean room which is sterilized as much as possible. Thus, if a cell is to be cultured in a large quantity for industrial application, there is a possibility that a burden on the workers is increased, time and costs are required for education/training of the workers, and human errors and mix-up of test substances and moreover, those carrying bacteria might cause contamination and the like, which requires much cost to cope with them. That makes a huge barrier in industrial application. Thus, by automating a series of culture work, those problems can be solved. 
     Regarding the automation of processes in the cell culture, an example of automation of a process to input a culture broth into a culture vessel such as cell seeding and medium replacement is described in Patent Document 1. In patent Document 1, by attaching/detaching a culture vessel and a joint on a manipulator side by a robot manipulator, a method of connecting the joint and the culture vessel is realized. At that time, leakage of culture broth from a tube of the joint and inside the culture vessel is prevented by a valve made of a resin film so that the connection can be made while a clean state is maintained. Also, the culture broth is supplied from beneath and discharged from above for broth replacement while the culture vessel connected by the manipulator stood in a vertical direction. 
     Also, a method for transferring a predetermined amount of a culture broth to a plurality of culture vessels more efficiently is proposed and its example is described in Patent Document 2. In this method, means for connecting a culture vessel and a flow passage by an aseptic connector, controlling communication of the broth by a three-way valve, and feeding the broth to the plurality of culture vessels branched in the flow passage is provided. 
     Patent Document 1: JP Patent Publication (Kokai) No. 2006-149237 
     Patent Document 2: JP Patent Publication (Kohyo) No. 63-503201 
     SUMMARY OF THE INVENTION 
     As methods of transplantation, there are methods of auto-transplantation in which a cell collected from himself/herself is in vitro treated and returned to himself/herself and allo-transplantation in which a cell collected from another person such as a donor body is in vitro treated for homotransplantations. In the auto-transplantation, since it is only necessary to create a necessary and sufficient amount of tissues for treating an affected portion of himself/herself and an amount of the cell that can be collected is small, culture is conducted using one or plural culture vessels. On the other hand, in the allo-transplantation, many cells provided from a donor body and the like are used and the number of cells is largely increased so as to produce numerous tissues, many culture vessels are used for culture. 
     Thus, if the culture vessels are treated one by one in processes of cell seeding or medium replacement of automatic culture for the allo-transplantation, a lot of time is required, which causes problems not only in the cost aspect but also non-uniform activation of the cells and medium and other environmental stresses might act on the cells and incur trouble in quality control. Thus, in order to reduce treatment time for the cell seeding and medium replacement, a method for feeding an equal amount of a culture broth to a plurality of culture vessels at the same time is required. 
     The present invention was made in view of such circumstances, and a fluid mechanism is provided that a plurality of culture vessels can be detachably attached to a flow passage at once, a broth is appropriately branched in the flow passage, and an equal amount of the culture broth is fed to the plurality of culture vessels. 
     In order to solve the above problems, the present invention provides the following means: 
     A culture vessel set that has one or plural recess portions in which one or plural culture vessels can be aligned and housed is provided. In the culture vessel set, m×n (row×column) pieces of culture vessels can be set in a matrix state in view of efficiency of space utilization. In the culture vessel set, spring-type fixtures are installed in a depth direction and a width direction, respectively, and the culture vessel side face is pressed by these two spring-type fixtures to be fixed to the culture vessel set and also made detachable. The culture vessel is configured to have a hole for supplying/discharging a culture broth from its upper face to/from the inside, and a manipulator having a joint for connecting the flow passage to the hole in the culture vessel is provided. 
     In the passage for feeding the broth, first, a cell is inputted from a cell input mechanism in an airlock structure, and cell concentration is adjusted in a tank. At that time, a process of cell input is removed in medium replacement. Using a pump, a cell suspension liquid is separated by a multi-way valve at the tank and fed to the culture vessel in the culture vessel set. At that time, at an outlet of the multi-way valve, n pieces of blocks having m pieces of outlets with an equal height in the horizontal direction are prepared for a single electromagnetic valve, and the liquid is put into the culture vessels at the equal height in the horizontal direction at the same time by the unit of block by ON/OFF of the electromagnetic valve. At that time, the electromagnetic valve in the outlet direction is controlled by a sensor on the outlet side so that an equal amount of medium feeding can be made, and the object can be realized. 
     That is, the cell culture device according to the present invention is provided with a liquid vessel for containing a liquid, a culture vessel set for holding a plurality of culture vessels provided with a liquid introduction port and a liquid exit port, respectively, liquid introduction/discharge means for introducing the liquid contained in the liquid vessel to the liquid introduction ports of the plurality of culture vessels held by the culture vessel set an discharging the liquid from the liquid exit ports of the culture vessels, a connecting member for connecting the plurality of liquid introduction ports and the plurality of liquid exit ports to the liquid introduction/discharge means, and state changing means for changing the culture vessel set from a horizontal state to a vertical state with its center line as a rotating axis when the liquid introduction/discharge means is operated. And when the culture vessel set is in the vertical state, the plurality of liquid exit ports and the plurality of liquid introduction ports are arranged with an interval between them in a perpendicular direction. 
     Here, the culture vessel set can hold m×n pieces (m is an integer of 2 or more, n is an integer of 1 or more) of culture vessels, and when the culture vessel set is in the vertical state, in m pieces of the culture vessels on the k-th stage (k is an integer of 1≦k≦n), a pressure of liquid feed by the liquid introduction/discharge means is the same at the plurality of liquid introduction ports, respectively. 
     The above-mentioned cell culture device is further provided with switch means for switching the liquid feed operation by the liquid introduction/discharge means to the plurality of culture vessels for m pieces of the culture vessels on the k-th stage. It may be so configured that a sensor for detecting discharge of the liquid from the exit port of m pieces of the culture vessels to which the liquid has been introduced by the liquid introduction/discharge means is provided, and when the sensor detects discharge of the liquid, the switch means switches the operation of the liquid introduction/discharge means so that the liquid is fed to m pieces of the culture vessels on the subsequent stage. 
     Also, the cell culture device according to the present invention is provided with a liquid vessel for containing a liquid, a culture vessel set for holding a plurality of culture vessels provided with a liquid introduction port and a liquid exit port, respectively, liquid introduction/discharge means for introducing the liquid contained in the liquid vessel to the liquid introduction ports of the plurality of culture vessels held by the culture vessel set and discharging the liquid from the plurality of liquid exit ports of the culture vessels, a connecting member for connecting the plurality of liquid introduction ports and the plurality of liquid exit ports to the liquid introduction/discharge means, and state changing means for changing the culture vessel set from a horizontal state to a vertical state when the liquid introduction/discharge means is operated. And when the culture vessel set is in the vertical state, the liquid introduction/discharge means feeds the liquid to the plurality of culture vessels located at the same height at the same time. 
     Also, the plurality of culture vessels are classified into a plurality of culture vessel groups to each of which the liquid is fed at the same time. In this case, the cell culture device is provided with switch means for switching the culture vessel group so that the liquid introduction/discharge means sequentially feeds the liquid to the plurality of culture vessel groups at the same time according to the switching operation by the switch means. 
     Additional features of the present invention will be made apparent from the best mode for carrying out the invention and the attached drawings. 
     According to the present invention, a plurality of culture vessels can be detachably attached to a flow passage at the same time, the liquid is branched appropriately by the flow passage, and an equal amount of a culture broth can be fed to the plurality of culture vessels. Thus, a cell can be cultured efficiently. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a diagram illustrating outline configuration of a cell culture device according to an embodiment of the present invention. 
         FIG. 2  is a diagram illustrating outline configuration of a culture vessel set that has one or plural recess portions inside and can fix the culture vessel by two spring-type fixtures. 
         FIG. 3A  is a conceptual diagram (1) relating to connection between a joint connected to a flow passage and the culture vessel set. 
         FIG. 3B  is a conceptual diagram (2) relating to connection between the joint connected to the flow passage and the culture vessel set. 
         FIG. 3C  is a conceptual diagram (3) relating to connection between the joint connected to the flow passage and the culture vessel set. 
         FIG. 3D  is a conceptual diagram (4) relating to connection between the joint connected to the flow passage and the culture vessel set. 
         FIG. 4  is a diagram illustrating a conceptual diagram relating to the flow passage (partially parallel type) when the joint and the culture vessel set are connected. 
         FIG. 5  is a flow-passage configuration diagram of multi-way valve in order to realize the flow passage. 
         FIG. 6  is an entire configuration diagram of the multi-way valve in the flow passage. 
         FIG. 7  is a detailed configuration diagram of a flow passage and a valve portion inside the multi-way valve. 
         FIG. 8  is a diagram illustrating block configuration of a liquid-feeding system according to the embodiment of the present invention. 
         FIG. 9  is a flowchart for explaining an operation of the liquid-feeding system according to the embodiment of the present invention. 
     
    
    
     DESCRIPTION OF SYMBOLS 
     
         
           1  cell culture device 
           10  culture vessel 
           11  culture space 
           12  connection hole 
           121  liquid injection port 
           122  liquid discharge port 
           13  culture vessel side face 
           14  culture chamber 
           15  culture broth with cell 
           16  air-lock chamber 
           161  outer door 
           162  inner door 
           163  flow-passage manipulator 
           17  pump 
           18  culture broth container 
           181  culture broth (for dilution) 
           19  tank 
           191  liquid level sensor 
           192  diluted culture broth with cell 
           20  culture vessel set 
           21  recess portion 
           22  spring-type fixture 
           23  removal hole 
           24  waste container 
           25  used culture broth (waste) 
           30  joint 
           31  flow passage 
           32  manipulator 
           33  rotating shaft 
           34  joint arm 
           35  motor 
           36  vertical direction 
           37  rotating direction 
           38  stopper 
           39  base 
           301  arm 
           302  motor 
           303  rotating shaft 
           304  connection groove 
           305  sensor 
           306  sensor 
           40  multi-way valve 
           41  electromagnetic valve 
           42  tube 
           43  connector 
           44  manifold 
           401  electromagnetic valve 
           402  electromagnetic valve 
           403  electromagnetic valve 
           411  spring 
           412  plug 
           413  flow passage 
           414  solenoid 
           415  electric wire 
           416  permanent magnet 
           417  diaphragm 
       
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     A preferred embodiment of the present invention will be described below referring to the attached drawings. However, attention should be paid that the present embodiment is only an example to realize the present invention and does not limit a technical scope of the present invention. In each drawing, the same reference numerals are given to common configuration. 
     &lt;Configuration of Cell Culture Device&gt; 
       FIG. 1  is a diagram illustrating outline configuration of a cell culture device  1  according to an embodiment of the present invention and particularly illustrates an entire liquid-feeding system inside a culture chamber  14  for injecting a liquid into a plurality of culture vessels  10 . 
     The cell culture device  1  is provided with the culture chamber  14 , an air-lock chamber  16  for input of a culture broth  15  with cell into the culture chamber  14 , and a waste container  24  for recovering a waste  25 . The air-lock chamber  16  is provided with an outer door  161  and an inner door  162  so that inside of the culture chamber  14  is not contaminated when the culture broth  15  with cell is inputted. Also, the cell culture device  1  is provided with, in the culture chamber  14 , a culture vessel set  20  for containing a plurality of culture vessels  10 , a joint  30  for injecting and discharging the culture broth  15  or a washing liquid with respect to each of the culture vessels  10 , a multi-way valve  40  for switching a flow of the culture broth  15 , a flow passage  31  (constituted by a plurality of tubes  42 ) provided between the joint  30  and the multi-way valve  40 , a culture broth container  18  for containing a culture broth  181  for dilution, a tank  19  connected to the air-lock chamber  16  and the culture broth container  18  and for diluting the culture broth  15  with cell, a pump  17  for moving the culture broth  15  with cell to the tank  19 , a liquid level sensor  104  for sensing a liquid coming from a waste port of the culture vessel  10 , an electromagnetic valve  105  for closing the tube  42 , and a controller  106  for controlling the electromagnetic valve  105  according to a sensing result by the liquid level sensor  104 . 
     &lt;Configuration of Culture Vessel and Culture Vessel Set&gt; 
       FIG. 2  is an outline diagram illustrating connection between the culture vessel  10  and the culture vessel set  20  according to this embodiment. 
     As shown in  FIG. 2 , the culture vessel set  20  has one or plural recess portions  21 , in which the culture vessel  10  is set. At that time, each culture vessel  10  is fixed in the recess portion  21  by pressing a culture vessel side face  13  in a P direction (See  FIG. 2 ) using two spring-type fixtures  22  in the recess portions  21 . The spring-type fixtures  22  are fixed to a bottom face or a side face of the recess portion  21  in the culture vessel set  20 . Also, the spring-type fixtures  22  are installed in the perpendicular direction to each other and use a material having strength such as stainless so that the fixture does not rust in a high-humidity environment during culture. When the culture vessel  10  is set in the recess portion  21 , the spring-type fixtures  22  press the culture vessel side faces  13  in the two directions (synthesized into a force in the P direction) so that a positional accuracy in a horizontal direction of the culture vessel  10  when the culture vessel  10  is set can be improved. Moreover, the force is applied on the whole culture vessel side faces  13  by that, movement in the perpendicular direction when an external force is applied or removal of the culture vessel  10  from the culture vessel set  20  can be prevented. For removal of the culture vessel  10 , a removal hole  23  provided at the culture vessel set  20  is used. 
     The culture vessel  10  has a culture space  11  for culturing cell in an internal closed system, in which a culture broth is placed and a cell is cultured. The culture vessel  10  has a pair of connection holes  12  for injection and discharge of the culture broth inside from the outside on an upper face portion of the culture vessel  10 . When the culture broth is injected, the culture broth is injected from one of the connection holes  12  (injection port), and liquid or air is discharged from the other connection hole  12  (discharge port). Since the culture vessel  10  has the connection holes  12  on the upper face portion, the culture vessels  10  set in the culture vessel set  20  all have the connection holes  12  arranged on the upper face portion of the culture vessel set  20 . Thus, by connecting the joint  30  connected to the flow passage  31  and the upper face portion of the culture vessel set  20 , the flow passage  31  and the culture vessels  10  can be connected to each other. 
     The culture vessel set  20  connected to the joint  30  is stood upright by a manipulator  32  from the horizontal direction to the perpendicular direction, the culture broth and the like is introduced from the connection hole (liquid injection port) located below of the two connection holes  12  and the liquid is injected. Thus, each culture vessel  10  is installed in the culture vessel set  20  so that injection of a diluted culture broth  192  into a liquid injection port  121 , which is the lower connection hole  12  of the culture vessel  10  and liquid or air discharge of a used culture broth  25  from a liquid discharge port  122 , which is the upper connection hole  12  of the culture vessel  10  can be carried out when all the culture vessels  10  installed in the culture vessel set  20  are stood upright. As mentioned above, by injecting the diluted culture broth  192  from below (liquid injection port) and discharging the used culture broth  25  or air from above (liquid discharge port), air bubbles having entered inside the culture space  11  are eliminated or the used culture broth  25  is discharged efficiently from above (liquid discharge port), and the uniformly diluted culture broth  192  can be injected into the culture space  11 . 
     &lt;Connection Between Joint and Culture Vessel Set&gt; 
       FIGS. 3A to 3D  are outline diagrams illustrating an operation relating to the connection between the joint  30  and the culture vessel set  20 , and using them the configuration of the manipulator  32  having the joint  30  for injecting liquid into the culture vessel  10  in the culture vessel set  20  and a series of operations of the connection between the culture vessel set  20  and the joint  30  by the manipulator will be described. 
       FIG. 3A  shows a state before the connection. First, configuration of each element will be described. The joint  30  is connected to the manipulator  32  by a rotating shaft  33  and a joint arm  34 . The joint arm  34  is moved vertically (in an arrow  36  direction) by a motor  35  of the manipulator  32 , and the joint  30  can be moved vertically (in the arrow  36  direction) with that motion. The joint  30  can be rotated from the horizontal direction to the perpendicular direction (in an arrow  37  direction) with the rotating shaft  33  as its axis. A stopper  38  is provided so that the joint  30  is not freely rotated at this time and the rotating shaft  33  can be locked and fixed at a predetermined position. A base  39  of the culture vessel set  20  is connected to an arm  301 . The arm  301  has a rotating shaft  303  connected to a motor  302 . Moreover, at the arm  301 , a connection groove  304  is provided so that the rotating shaft  303  and the rotating shaft  33  of the joint  30  are overlapped on the same line. 
     When the culture vessel set  20  is inserted into the base  39  as shown in  FIG. 3B , a sensor  305  senses presence of the culture vessel set  20 . Next, the motor  35  lowers the joint  30  and moves the rotating shaft  33  of the joint  30  to the connection groove  304 . When a sensor  306  in the connection groove  304  senses that the rotating shaft  33  is inside the connection groove  304 , control is executed (by a motor control portion, not shown) so that the operation of the motor  35  is stopped. 
     When the sensor  306  in the connection groove  304  senses the presence of the rotating shaft  33  of the joint  30 , the joint  30  and the culture vessel  10  are connected to each other as shown in  FIG. 3C , and each rotating shaft (the rotating shaft  33  and the rotating shaft  303 ) are present on the same line. When the rotating shaft  303  of the motor  302  connected to the arm  301  is rotated (in the arrow  37  direction) in this state, the rotating shaft  33  of the joint  30  is similarly rotated (in the arrow  37  direction), and the culture vessel set  20  on the base  39  and the joint  30  can be rotated (in the arrow  37  direction) in a state connected to each other. 
       FIG. 3D  shows a state where the joint  30  and the culture vessels  10  are rotated from the horizontal direction to the perpendicular direction (in the arrow  37  direction), and the joint  30  and the culture vessel set  20  are stood in the vertical state. In this state, the (diluted) culture broth flows into the flow passage  31  to be injected into the culture vessel  10 . At that time, it is configured such that the culture broth is injected from the liquid injection port  121 , which is the lower connection hole  12  of the culture vessel  10 , while waste liquid or air flows out of the liquid discharge port  122 , which is the upper connection hole  12  of the culture vessel  10  forming the pair. 
     After the culture vessel has been injected, the state is returned to the horizontal to that in  FIG. 3B , the joint  30  is removed from the culture vessel  10  into a completely separated state as in  FIG. 3A , and replacement of the culture broth or injection of cells in the culture broth is finished. 
     &lt;Flow-Passage Configuration for Liquid Injection and Discharge&gt; 
     Next, a flow-passage configuration for injecting the culture broth into the plurality of culture vessels  10  installed in the culture vessel set  20  will be described. First, merits and demerits of each of the most basic types of serial type and parallel type will be described. 
     (1) In the flow-passage configuration of the serial type, the liquid is put into one single culture vessel  10 , its liquid discharge port  122  is connected to the liquid injection port  121  of the subsequent culture vessel  10 , and all the culture vessels  10  are connected. This serial-type configuration has a merit that the flow passage and a liquid-feed system is simple, but it has demerits of non-uniformity of the culture broth that the culture broth in the last culture vessel  10  is not fresh at medium displacement, a pressure difference caused by a height when being stood makes control difficult, and contamination by fungi or bacteria of one culture vessel  10  contaminates all. 
     On the other hand, in the parallel flow-passage configuration, the liquid is injected to all the culture vessels  10  independently in parallel. The parallel-type configuration has merits that even if one culture vessel  10  is contaminated by fungi or bacteria, the other culture vessels  10  are hardly contaminated, the culture broth can be injected uniformly since the culture broth is injected independently, and a pressure difference is not caused at the injection between the culture vessels  10  due to the heights of the culture vessels  10 , but this type also has a demerit that the flow passage and the liquid-feeding system might become extremely complicated. 
     The inventors have conceived a method utilizing the respective merits (this embodiment) based on the examination results. That is, it is the partially parallel-type flow-passage configuration that when the culture vessel set  20  connected to the joint  30  is stood in the vertical direction, the culture vessels  10  aligned in the horizontal direction are connected in parallel and the liquid is fed for each unit. 
     (2) Referring to  FIG. 4 , configuration of the partially parallel-type flow passage  31  and the configuration of the joint  30  to be connected to the culture vessel set  20  will be described. 
     As shown in  FIG. 4A , it is configured that when the culture vessel set  20  connected to the joint  30  is rotated in the arrow  37  direction and stood in the vertical direction, parallel treatment can be conducted with the lower connection holes  12  of the culture vessels  10  aligned in the horizontal direction as the liquid injection ports  121  and the upper connection holes  12  as the liquid discharge ports  122 . That is, as shown in  FIG. 4B , the parallel treatment is conducted by the unit of each group of  10 - 1 ,  10 - 2 ,  10 - 3 . 
     By additionally arranging the plurality of culture vessels  10  aligned horizontally in the perpendicular direction in the state where the culture vessel set  20  is stood upright, the merit of the serial type can be obtained that the flow passage and the liquid-feeding system are simplified and the merit of the parallel type such as difficulty in contamination of the whole, uniform injection of the culture broth, no pressure difference by the heights of the culture vessels  10  and the like can be both obtained in the configuration. 
     &lt;Operation of Parallel-Type Liquid-Feeding/Liquid-Discharge System&gt; 
     Returning to  FIG. 1 , a flow of the culture broth in the liquid injection/discharge system having an entire flow passage to realize the partially parallel flow passage  31  will be described. 
     The culture broth  15  with cell is inputted from the outer door  161  of the air-lock chamber  16  installed at the cell culture device  1  and enters the air-lock chamber  16 . Then, the inner door  162  of the air-lock chamber  16  is opened, a flow-passage manipulator  163  is lowered by motor driving from there, and the culture broth  15  with cell is sucked by the pump  17  and moved to the tank  19 . Also, the culture broth  181  for dilution is moved to the tank  19  by the pump  17  and density of the cell is adjusted. At that time, the culture broth  192  in the tank  19  is monitored by a tank liquid level sensor  191 . Here, at medium replacement, the liquid-feeding process of the culture broth  15  for cell is not required, but only the culture broth  181  for dilution is moved to the tank  19 . 
     And the diluted culture broth  192  is fed by the pump  17  to the multi-way valve  40  from the tank  19 . After that, the diluted culture broth  192  is branched by the multi-way valve  40  made of an electromagnetic valve  41 , and the diluted culture broth  192  is fed into the culture vessel  10 . Here, the liquid is fed only to the culture vessels  10  (culture vessel group  10 - 1 ) having a height y 1  in the perpendicular direction to the liquid injection port  121  of the culture vessel  10  from the multi-way valve  40  at the same time. The liquid is similarly fed to y 1 +y 2  (culture vessel group  10 - 2 ), y 1 +y 2 +y 3  (culture vessel group  10 - 3 ) in  FIG. 1 . Here, diameters of the tube  42  and a connector  43  used by each group are set the same so that a difference by pressure is not caused other than the height. In the same group, since the liquid-injection positions and the liquid discharge positions (height) are set the same, it is configured such that a pressure difference is not caused at the liquid injection and the liquid discharge. 
     The extra culture broth coming out of the liquid discharge port  122  of the culture vessel  10  is sensed by the liquid level sensor  104  as the used culture broth  25 . On the basis of the sensing result, the controller  106  closes the tube  42  by a pinch-valve like electromagnetic valve  105 . Thereby, the diluted culture broth  192  can be easily put into the other culture vessels  10  to which the liquid is fed at the same time. Here, when the electromagnetic valve  105  of the same liquid-feeding system is closed (when the extra culture broth comes out of the liquid discharge ports of all the culture vessels in the same group and the electromagnetic valve  105  is closed), the electromagnetic valve  41  of the multi-way valve  40  is switched to another valve and the liquid is fed to the culture vessels  10  with another height (another culture vessel group). At that time, if all the electromagnetic valves  105  of the same liquid-feeding system are not closed but such an amount of the culture broth  192  that can fill all the culture vessels  10  in the culture vessel group is being fed, an error is displayed as liquid leakage. When the electromagnetic valves.  105  of all the culture vessel groups are closed, it is considered as work finished, the joint  30  and the culture vessel set  20  are removed, and the culture is continued. The waste liquid remaining in the flow passage  31  is all moved to the waste container  24 . 
     &lt;Configuration of Multi-Way Valve&gt; 
     Using  FIGS. 5 to 7 , configuration of the multi-way valve  40  will be described. The multi-way valve  40  in this embodiment has one port of inlet and nine ports of outlet. The culture vessel set  20  is configured such that laterally three vessels and vertically three vessels, totaling in nine culture vessels  10  can be set. In the multi-way valve  40 , the number of ports can be increased to  20  at the maximum. 
       FIG. 5  shows a fluid circuit. When the culture vessel set  20  is stood in the vertical direction (See  FIG. 3D ), since the number of culture vessels included in the same culture vessel group in the horizontal direction is  3 , in the multi-way valve  40 , one electromagnetic valve  41  has parallel three ports and since there are three rows in the perpendicular direction, three sets of the electromagnetic valves  41  having the parallel three ports are provided, totaling in nine ports. At a port number A, the culture broth  192  is put into the multi-way valve  40 , while port numbers B, C, and D are parallel at an electromagnetic valve SV 1  ( 401 ), port numbers E, F, and G at an electromagnetic valve SV 2  ( 402 ), and port numbers H, I, and J at an electromagnetic valve SV 3  ( 403 ) so that the culture broth  192  can be discharged in parallel. 
       FIG. 6  is a view illustrating arrangement and configuration of each port and each electromagnetic valve  41  in a three-dimensional manner.  FIG. 7A  is a top view of the multi-way valve  40 , and  FIG. 7B  is a view illustrating inner configuration of each electromagnetic valve  41 . 
     The multi-way valve  40  is constituted by a manifold  44  having a flow passage  413  and the electromagnetic valve  41  of a diaphragm type  417 . This electromagnetic valve  41  is in a normally closed (NC) state where a plug  412  is pressed by a spring  411  onto the flow passage  413 , but when electricity from a wire  415  flows through a solenoid  414  so as to form a magnetic field, a permanent magnet  416  is lifted and the plug  412  is removed, by which the flow passage  413  is opened. When the electricity is stopped, the permanent magnet  416  is lowered by the spring  411  and the plug  412  is placed. The tube  42  from the outside and the manifold  44  are connected to each other by the connector  43 . The culture broth  192  from the tank  19  first enters the manifold  44  through A, but since all the electromagnetic valves  41  are closed, the liquid does not flow. When the electricity flows from the controller and the electromagnetic valve SV 1  ( 401 ) is opened, for example, equal amounts of the culture broth  192  flow from the connectors B, C, and D. Here, in order to eliminate the pressure difference caused by a difference in the heights, the heights from the manifold  44  bottom face to the connector  43  are all equal. 
     &lt;Software Realization of Liquid-Feeding Operation&gt; 
     In the present invention, the above-described liquid-feeding operation can be realized using a computer program.  FIG. 8  is a block diagram illustrating configuration of a portion relating to computer control in the entire liquid-feeding system by the embodiment of the present invention. Each block in  FIG. 8  is connected by a bus  100 .  FIG. 9  is a flowchart for explaining the operation of the liquid-feeding system.
     (1) In  FIG. 8 , a CPU  101  controls the entire operation of the liquid-feeding system and constitutes at least a control entity of the operation by the flowchart shown in  FIG. 9 . That is, the CPU  101  reads out a control program based on the flowchart in  FIG. 9  stored in a ROM  102  upon operation start of the system, extends it on a RAM  103  and controls the entire system. It is possible that the operation of the controller  106  is executed by the CPU  101 .   

     The pump  17  is used for moving the culture broth in the flow passage. The pump  17  is operated forward or backward or its speed can be changed by the CPU  101 . The tank liquid level sensor  191  senses that a liquid is filled in the tank  19  with light in a band not damaging a cell and the like and transmits a signal to a control portion. The waste liquid level sensor  104  is connected to the tube  42  so as to sense if the liquid is filled in the tube  42  and transmit a signal to the controller  106 . The sensor is different depending on whether it is for cell seeding without culture broth in the culture vessel  10  in the beginning or it is for medium replacement with the culture broth in the beginning. In the multi-way valve electromagnetic valve  41  and the electromagnetic valve  105 , the valves are opened/closed by flowing electricity to the solenoid upon the signal from the respective sensors. 
     An input portion  107  is for a user to instruct carrying-in/-out of the culture vessel, for example, and corresponds to a keyboard, a mouse and the like. A display portion  108  displays that the culture system is in operation, instruction contents from the user and the like and corresponds to a CRT, a liquid crystal display and the like.
     (2) Subsequently, the operation of the liquid-feeding system provided with the configuration as in  FIG. 8  will be described using  FIG. 9 . Here, the medium replacement mainly operating in the liquid-feeding system will be described. The culture broth  15  containing cell is fed to the tank  19  by the pump  17  before this process. Unless otherwise specified, the operation entity is the CPU  101 .   

     When the medium replacement is required (Step S 901 ), the pump  17  is operated so as to feed the culture broth  181  to the tank  19  (Step S 902 ). The sensor  191  senses that the culture broth  181  is filled in the tank  19  (Step S 903 ), and an amount of the culture broth  192  is determined (Step S 904 ). When a predetermined amount of the culture broth  192  is filled, processing moves to Step S 905 , and if the culture broth  192  is not completely filled, the routine returns to Step S 903 , and the pump  17  is continuously operated. Here, the liquid level of the culture broth  192  is detected by the liquid level sensor  191 , and a detection result is transmitted to the CPU  101 . 
     If it is determined at Step S 904  that the entire culture broth  192  is filled, the operation of the pump  17  is stopped (Step S 905 ). Electricity is flown to the electromagnetic valve  41  of the multi-way valve  40  required for liquid-feeding so as to open the flow passage  413  (Step S 906 ) and to operate the pump  17 , and the culture broth  192  in the tank  19  is fed into the culture vessel  10  (Step S 907 ). Connection between the culture vessel set  20  and the joint  30  has been completed so far, and the culture vessel set  20  is in a standing state as in  FIG. 3D . 
     The culture broth  192  is filled in the culture space  11  inside the culture vessel  10 , the extra culture broth or the used culture broth  25  comes out of the liquid discharge port  122  through the tube  42 , and if the waste liquid sensor  104  senses that (Step S 908 ), electricity is made to flow to the electromagnetic valve  105  in the tube  42  (Step S 909 ), and discharge of the extra culture broth is stopped by closing the flow passage  413 . In the culture vessel  10  placed in parallel (the same culture vessel group), upon sensing by the waste liquid sensor  104  of the tube  42  coming out of the liquid discharge port  122 , if electricity does not flow through the electromagnetic valve  105 , the routine returns to Step S 908 , and the pump  17  is continuously operated so as to continue flowing the diluted culture broth  192 . However, even if all the electromagnetic valves  105  are not operated after an amount that can be replaced by the entire diluted culture broth  192  is flown, it is considered to be caused by liquid leakage or a failure in the device, and an error code is displayed on the display portion. If it is determined at Step S 910  that all the electromagnetic valves  105  are being operated, the routine goes on to Step S 911 , the pump  17  is stopped, and the electromagnetic valve  41  of the multi-way valve  40  is closed (Step S 911 ). If all the electromagnetic valves  41  of the multi-way valve  40  are not operated, the electromagnetic valve  41  not in operation is opened (Step S 914 ), and the routine goes to Step S 907 . If it is determined at Step S 912  that all the electromagnetic valves  41  are being operated, the routine goes on to Step S 913 , and the liquid-feeding work is finished. 
     The function of the embodiment can be also realized by a program code of the realizing software. In this case, a recording medium in which the program code is recorded is provided to the system or the device, and the system or a computer of the device (or CPU or MPU) reads out the program code stored in the recording medium. In this case, the program code itself read out of the recording medium realizes the above-mentioned function of the embodiment, and the program code itself and the recording medium storing it constitute the present invention. The recording mediums providing the program code include a floppy (registered trademark) disk, CD-ROM, DVD-ROM, hard disk, optical disk, magnetooptical disk, CD-R, magnetic tape, non-volatile memory card, ROM and the like. 
     Also, on the basis of an instruction of the program code, OS (operating system) operating on the computer may execute a part of or the whole of the actual processing so that the above-mentioned function of the embodiment is realized by the processing. Moreover, after the program code read out of the recording medium is written in a memory on the computer, on the basis of the instruction of the program code, a part of or the whole of the actual processing may be executed by a CPU of the computer and the like so that the above-mentioned function of the embodiment is realized by the processing. 
     Also, it may be so configured that a program code of the software that realizes the function of the embodiment is delivered through a network, the code is stored in storage means such as a hard disk of the system or the device, a memory and the like or a recording medium such as CD-RW, CD-R and the like and the computer (or CPU or MPU) of the system or the device reads out and executes the program code stored in the storage means or the recording medium. 
     &lt;Conclusion&gt; 
     In the flow-passage structure by the embodiment of the present invention, a single electromagnetic valve is allocated to all the culture vessels at an equal height in the horizontal direction, and an equal amount of a liquid is fed at the same time by controlling ON/OFF of the electromagnetic valve. Thereby, the liquid can be fed to the plurality of cartridges at the same time. Thus, efficiency of the entire flow-passage system and reduction of the liquid-feeding time can be promoted, by which damage applied to a cell such as an external force by a temperature and water pressure can be minimized. 
     Here, the culture vessels to which the liquid is fed at the same time and the outlet from the electromagnetic valve are set at the equal height in the horizontal direction because a difference in a medium injection amount caused by water head difference by the height at the height of the culture vessel according to Pascal&#39;s principle is to be eliminated. At this time, a diameter and a length of a flow passage from the electromagnetic valve to the culture vessel are supposed to be all the same. Thereby, a pressure to each culture vessel at the liquid-feeding can be made equal. 
     Also, the culture vessel and the joint can be detached. Thus, the inside of the flow passage can be made washable so that clogging in the flow passage caused by solidification of protein and the like can be eliminated. 
     Moreover, if the culture vessel and the joint are not connected, it is a closed-system culture vessel, while if the culture vessel and the joint are connected, the flow passage is made through. Thus, bacteria or fungus does not intrude but the liquid can be fed in a clean state. 
     Alternatively, a sensor may be provided in the flow passage after the liquid-feeding so that the flow passage is blocked by an electromagnetic valve on the outlet side on the basis of a detection result of the sensor. Thereby, a required amount of medium can be reduced, and efficiency can be improved. 
     Though not described in the embodiment, in the present invention, the shapes of the culture vessel set and the joint can be changed. Thereby, it is possible to respond to various shapes of the culture vessels or one or plural culture vessels. Therefore, even if a culture vessel is different depending on a tissue to be cultured, the system can be used, and an extremely flexible cell culture system can be constructed. 
     The present invention can respond to various culture conditions by the culture vessel set or the multi-way electromagnetic valve to one or plural culture vessels as mentioned above or moreover, only by changing the shapes of the joint and the culture vessel set. Thereby, in the automation of the cell culture work requiring high cleanliness realized by an unattended room, general-purpose properties can be provided, numerous culture vessels can be coped with, and medium can be injected with high efficiency and uniformity, and thus, demand is high in the industries including cell transplantation and regeneration medicine and availability is expected to be high.

Technology Classification (CPC): 2