Patent Publication Number: US-2019185802-A1

Title: Cell culturing device, cell culturing system and cell culturing method

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     The present patent application is a Divisional of U.S. patent application Ser. No. 15/302,707, filed Oct. 7, 2016, which is a National Phase of International application No. PCT/JP2014/060572, filed Apr. 14, 2014, all of which are incorporated herein by reference in their entireties. 
    
    
     TECHNICAL FIELD 
     The present invention relates to a cell culturing device, a cell culturing system and a cell culturing method. 
     BACKGROUND ART 
     Many studies on an artificial liver and the like have been conducted. However, cell culture of only a hepatic parenchymal cell for a long period of time has not been realized. The present inventors have reported that high liver functions are expressed by using a co-culture system of an endothelial cell and a hepatic parenchymal cell which forms a structured network due to the selection of a scaffold material, in the short term (see Non-Patent Document 1). A structure of a device suited for this system is disclosed, for example, in Patent Document 1. 
     PRIOR ART DOCUMENTS 
     Patent Document 
     
         
         Patent Document 1: Japanese Patent Laid-open Publication No. 2011-244713 
       
    
     Non-Patent Document 
     
         
         Non-Patent Document 1: Yoichi Fujiyama, Yoh-ichi Tagawa, and 5 other persons, “Analysis of liver functions in a co-culture with a hepatic parenchymal cell by using a system for culturing tubular endothelial liver cells”, Proceedings of The 30th Annual Meeting of The Molecular Biology Society of Japan, 2007 
       
    
     SUMMARY OF THE INVENTION 
     Problems to be Solved by the Invention 
     In the evaluation of pharmacokinetics, various methods have been studied in order to utilize an evaluation in a liver tissue. In addition, a device and a system for culturing a cell which use a co-culture system have also been developed. 
     However, a drug is not always directly incorporated into the liver in vivo, and therefore, it is preferable to consider the influence of the absorption of an ingredient in an intestine in the case of oral administration. 
     An object of the present invention is to provide a cell culturing device which is capable of culturing two types of cells and is capable of introducing a metabolite of one type of cells into the other type of cells, as well as a cell culturing system and a cell culturing method which utilize the cell culturing device. 
     Solutions to the Problems 
     The cell culturing device according to the present invention has: a first culture chamber; a first introduction flow channel and a first discharge flow channel which are connected to the first culture chamber; a second culture chamber which is connected to a halfway part of the first introduction flow channel via a first porous membrane; and a second introduction flow channel and a second discharge flow channel which are connected to the second culture chamber. 
     The cell culturing system according to the present invention has: the cell culturing device according to the present invention; and a culture medium feeding part which feeds a culture medium into the first culture chamber by connecting a pump to the cell culturing device. 
     The cell culturing method according to the present invention has the steps of: culturing cells in the first culture chamber and in the second culture chamber, respectively, by using the cell culturing device according to the present invention; and introducing a liquid in the second culture chamber into the first culture chamber via the first porous membrane and the first introduction flow channel. 
     Effects of the Invention 
     The cell culturing device, the cell culturing system and the cell culturing method according to the present invention are capable of culturing two types of cells and are capable of introducing a metabolite of one type of cells to the other type of cells. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a schematic plan view and a cross-sectional schematic diagram for illustrating an example of a cell culturing device. 
         FIG. 2  is a plan view which shows the cell culturing device according to the same example by disassembling the cell culturing device. 
         FIG. 3  is a schematic diagram for illustrating an example of a cell culturing system. 
         FIG. 4  is a schematic diagram for illustrating another example of a cell culturing system. 
         FIG. 5  is a schematic diagram for illustrating a further example of a cell culturing system. 
         FIG. 6  is a schematic diagram for illustrating a further example of a cell culturing system. 
         FIG. 7  is a schematic diagram for illustrating a further example of a cell culturing system. 
         FIG. 8  is a schematic diagram for illustrating a further example of a cell culturing system. 
         FIG. 9  is a schematic plan view and a cross-sectional schematic diagram for illustrating another example of a cell culturing device. 
         FIG. 10  is a plan view which shows the cell culturing device shown in  FIG. 9  by disassembling the cell culturing device. 
     
    
    
     EMBODIMENTS OF THE INVENTION 
     An example of the cell culturing device according to the present invention includes a cell culturing device in which the first discharge flow channel is connected to the first culture chamber via a second porous membrane. By this, for example, the clogging of the first discharge flow channel due to the influx of a substance in the first culture chamber such as a cell and a scaffold material (a gel) into the first discharge flow channel can be prevented. However, it is not necessary to arrange the second porous membrane in the cell culturing device according to the present invention. 
     An example of the cell culturing device according to the present invention includes a cell culturing device, in which the first introduction flow channel has a liquid collecting part between the first culture chamber and the second culture chamber, and in which at least a part of the wall of the liquid collecting part is formed by an elastic member which is capable of being penetrated by a suction implement having a sharp tip, wherein the through hole is capable of being closed when the suction implement is withdrawn after the penetration due to the elasticity of the member. By this, a liquid which has passed through the second culture chamber, for example, a liquid which contains a metabolite of cells cultured in the second culture chamber, can be collected before the liquid is introduced into the first culture chamber. However, it is not necessary for the first introduction flow channel to have the liquid collecting part, in the cell culturing device according to the present invention. 
     An example of the cell culturing device according to the present invention includes a cell culturing device which has a main body part and a lid part which is detachably attached to the main body part, in which the first culture chamber is formed in the main body part, and has an opening on the surface of the main body part which is in contact with the lid part. By this, in a state where the main body part and the lid part are separated, for example, a substance such as a cell, a scaffold material and a culture medium can be placed in the first culture chamber without interposing the first introduction flow channel and the first discharge flow channel. However, the cell culturing device according to the present invention may have a structure which does not have the detachable main body part or the lid part. 
     In addition, an example of the cell culturing device includes a cell culturing device in which the material of a contact surface between the main body part and the lid part is a PDMS (polydimethylsiloxane) or a silicone rubber. By this, the lid part can be stably fixed to the main body part due to the self-adsorption property, for example, without using a special fixing means such as a screw and an adhesive agent. In addition, the main body part can be easily attached/detached to/from the lid part, as needed. However, the material of a contact surface between the main body part and the lid part is not limited to a PDMS or a silicone rubber, and materials other than the PDMS and the silicone rubber may be employed. 
     Further, an example of the cell culturing device includes a cell culturing device, in which at least one of the surface of the main body part in contact with the lid part and the surface of the lid part in contact with the main body part is formed in a convex shape, and in which the plane size of the contact surface between the main body part and the lid part is smaller than the plane size of the device. By making the plane size of the contact surface between the main body part and the lid part smaller, a force distribution to an unnecessary part can be suppressed, and therefore, the adherence between the main body part and the lid part can be improved. However, both the surface of the main body part in contact with the lid part and the surface of the lid part in contact with the main body part may be flat surfaces, in the cell culturing device according to the present invention. 
     Furthermore, an example of the cell culturing device includes a cell culturing device in which the second culture chamber is formed in the lid part. By this, cells can be cultured in the first culture chamber and in the second culture chamber, respectively, under conditions and environments different from each other, by separating the main body part in which the first culture chamber is formed and the lid part in which the second culture chamber is formed. However, the second culture chamber may be formed in the main body part, or the second culture chamber may also be formed to be astride the main body part and the lid part, in the cell culturing device according to the present invention. 
     An example of the cell culturing device according to the present invention includes a cell culturing device, in which a co-culture system containing at least an endothelial cell and a hepatic parenchymal cell is co-cultured in the first culture chamber, and in which a cell derived from an intestine is cultured in the second culture chamber. By this, a metabolite of the cell derived from the intestine can be fed to the co-culture system which contains the endothelial cell and the hepatic parenchymal cell, and therefore, an environment closer to in vivo can be realized. However, the cell cultured in the first culture chamber and that cultured in the second culture chamber are not limited to these cells, in the cell culturing device according to the present invention. 
     In addition, an example of the cell culturing device according to the present invention includes a cell culturing device, in which a gel which serves as a cell scaffold material is housed in the first culture chamber, and in which the co-culture system having a tubular structure forms a structured network on the gel. 
     An example of the cell culturing system according to the present invention includes a cell culturing system, in which the culture medium feeding part feeds the culture medium to the first culture chamber by circulating the culture medium, and in which the cell culturing system further has a dialysis part in a halfway part of the culture medium circulating line of the culture medium feeding part. By this, the culture medium can be fed into the first culture chamber in a circulated manner while removing unnecessary substances by the dialysis part. This constitution is particularly useful in the case where a co-culture system, which contains at least an endothelial cell and a hepatic parenchymal cell, is co-cultured in the first culture chamber as well as a cell derived from an intestine is cultured in the second culture chamber. However, it is not necessary for the cell culturing system according to the present invention to have the dialysis part. 
     An example of the cell culturing method according to the present invention includes a cell culturing method, in which cells are cultured in the main body part of the first culture chamber and in the second culture chamber which is formed in the lid part, respectively, in a state where the main body part and the lid part are separated from each other, by using the cell culturing device according to the present invention in which the second culture chamber is formed in the lid part, and thereafter, the main body part and the lid part are joined together. By this, cells can be cultured in the first culture chamber and in the second culture chamber, respectively, under conditions and environments different from each other. However, in the cell culturing method according to the present invention, cells may be cultured in the first culture chamber and in the second culture chamber, respectively, in a state where the main body part and the lid part of the cell culturing device are joined together. 
     An example of the cell culturing method according to the present invention includes a cell culturing method, in which a co-culture system containing at least an endothelial cell and a hepatic parenchymal cell is co-cultured in the first culture chamber, and in which a cell derived from an intestine is cultured in the second culture chamber. By this, a metabolite of the cell derived from the intestine can be fed to the co-culture system which contains the endothelial cell and the hepatic parenchymal cell, and therefore, an environment closer to in vivo can be realized. However, the cell cultured in the first culture chamber and that cultured in the second culture chamber are not limited to these cells, in the cell culturing method according to the present invention. 
     Further, an example of the cell culturing method according to the present invention includes a cell culturing method, in which a gel which serves as a cell scaffold material is housed in the first culture chamber, and in which the co-culture system having a tubular structure is caused to form a structured network on the gel. 
     Furthermore, an example of the cell culturing method according to the present invention includes a cell culturing method, in which a reagent is introduced into the second culture chamber in which the cell derived from the intestine is cultured, from the second introduction flow channel, and a metabolite of the cell derived from the intestine thus obtained is introduced into the first culture chamber via the first porous membrane and the first introduction flow channel. 
     In addition, an example of the cell culturing method according to the present invention includes a cell culturing method in which the culture medium discharged from the first culture chamber is dialyzed and is fed into the first culture chamber in a circulated manner. By this, an environment closer to in vivo can be realized. 
     The present invention is a technique for culturing cells, and for example, is a useful technique which is used for an artificial organ such as an artificial liver, or a drug metabolism test and the like. 
     The cell culturing device according to the present invention has, for example, a second culture chamber which is connected via a first porous membrane to a halfway part of a flow channel (a first introduction flow channel) of a culture solution which is to be introduced into a first culture chamber in which a liver tissue is cultured. For example, a cell derived from an intestine is cultured in the second culture chamber and a drug to be tested is administered into the culture medium of the second culture chamber. 
       FIG. 1  is a schematic plan view and a cross-sectional schematic diagram for illustrating an example of a cell culturing device.  FIG. 2  is a plan view which shows the cell culturing device according to this example by disassembling the cell culturing device. 
     A cell culturing device  1  according to this example is roughly formed by a main body part  3  and a lid part  5 . The main body part  3  is, for example, a part for culturing a liver tissue. The lid part  5  is, for example, a part for culturing an intestinal epithelial cell. The lid part  5  is detachable/attachable from/to the main body part  3 . 
     The main body part  3  has a base plate  7 , a PDMS block  9  and a PDMS block  11 . 
     The base plate  7  of the main body part  3  is, for example, formed of synthetic quartz. Meanwhile, the base plate  7  is not limited to that having a plane shape. The base plate  7  may be anything as long as it is capable of being adsorbed by the PDMS block  9 . For example, the base plate  7  may be a culture dish. 
     The PDMS block  9  and the PDMS block  11  are, for example, formed of SILPOT 184 (manufactured by Dow Corning Toray Co., Ltd.). The plane size of the PDMS blocks  9  and  11  is, for example, 25 mm×25 mm (millimeter). The thickness of the PDMS block  9  is, for example, 2.0 mm. The thickness of the PDMS block  11  is, for example, 1.0 mm. 
     The PDMS block  9  has a through hole  9   a  which has a diameter of, for example, 10 mm. 
     The PDMS block  11  has a through hole  11   a , a recessed part lib, a protruding part  11   c  and a through hole  11   d.    
     The through hole  11   a  is formed at the position which is overlapped with the through hole  9   a , when the PDMS block  9  and the PDMS block  11  are stuck together. The through hole  11   a  has, for example, a diameter of 10 mm. 
     The recessed part lib is formd on the surface in contact with the PDMS block  9 . The depth of the recessed part lib is, for example, about 0.2 mm. The recessed part  11   b  has a peripheral part around the through hole  11   a  and a projected part protruding from the peripheral part. The width of the recessed part lib at the peripheral part around through hole  11   a  is, for example, about 1.0 mm. With regard to the size of the projected part in the recessed part  11   b , for example, the width is about 2.0 mm and the length is about 2.0 mm. 
     The protruding part  11   c  is formd in a convex shape bulging from the surface which is opposed to the surface in contact with the PDMS block  9 . The height of the protruding part  11   c  is, for example, about 0.5 mm. The upper surface of the protruding part  11   c  constitutes the surface in contact with the lid part  5 . The protruding part  11   c  is formed at the position which contains the position where the through hole  11   a  is formd. Meanwhile, the thickness of the PDMS block  11  is the thickness including the thickness of the protruding part  11   c.    
     The through hole  11   d  is formed at the position which is overlapped with the projected part in the recessed part lib. The diameter of the through hole  11   d  is, for example, 1.5 mm. 
     The forming process of the main body part  3  is briefly explained. 
     The PDMS block  11  in which the recessed part lib and the protruding part  11   c  are formd is prepared. The recessed part lib has a circular part having a diameter of about 12 mm and a projected part protruding from the circular part. The through hole  11   d  having a diameter of 1.5 mm is opened at the position which is overlapped with the projected part. 
     The PDMS block  11  in which the through hole  11   a  has not been formed and the PDMS block  9  in which the through hole  9   a  has not been formd are stuck together. At this time, it is preferable that the contact surfaces between the PDMS block  9  and the PDMS block  11  are subjected to an oxygen plasma treatment before being stuck together. The surface of the PDMS block  11  in contact with the PDMS block  9  is the surface on which the recessed part lib is formed. 
     A through hole having a diameter of 10 mm and passing through the PDMS blocks  9  and  11  which are stuck together is formed so that the through hole is overlapped with the center of the circular part in the recessed part lib. By this, the through hole  9   a  is formed in the PDMS block  9 , and the through hole  11   a  is formed in the PDMS block  11 . 
     The surface of the PDMS block  9  which is opposed to the PDMS block  11  and the base plate  7  are stuck together so that the main body part  3  is completed. At this time, it is preferable that the surface of the PDMS block  9  in contact with the base plate  7  is subjected to an oxygen plasma treatment. Meanwhile, it is also possible to stick the PDMS block  9  onto the base plate  7 , a cell culture dish, and the like by utilizing the self-adsorption property of the PDMS block  9  though the oxygen plasma treatment is not performed. 
     Next, the lid part  5  is explained. 
     The lid part  5  has a silicone rubber sheet  13 , a filter  15  (a first porous membrane), a filter  17  (a second porous membrane), a silicone rubber sheet  19 , a PDMS block  21 , a silicone rubber sheet  23 , a PDMS block  25  and a PDMS block  27 . 
     The plane size of the silicone rubber sheets  13 ,  19  and  23  as well as the PDMS blocks  21 ,  25  and  27  is, for example, 25 mm×25 mm. 
     The silicone rubber sheets  13 ,  19  and  23  are, for example, formed of a Silius extremely thin silicone rubber sheet (a product of Fuso Rubber Co., Ltd). 
     The PDMS blocks  21 ,  25  and  27  are, for example, formed of SILPOT 184 (manufactured by Dow Corning Toray Co., Ltd.). 
     The silicone rubber sheet  13  and the silicone rubber sheet  19  are provided in order to hold the filters  15  and  17  by sandwiching the filters  15  and  17 . The thickness of each of the silicone rubber sheets  13  and  19  is, for example, 0.1 mm. 
     The silicone rubber sheet  13  has a long hole  13   a , a through hole  13   b , a long hole  13   c , a through hole  13   d  and a through hole  13   e , which are, for example, formd by a method such as a punching method. 
     The width dimension of the long hole  13   a  is, for example, 2.0 mm. One end of the long hole  13   a  is connected to the through hole  13   b.    
     The diameter of the through hole  13   b  is, for example, 4.0 mm. 
     The width dimension of the long hole  13   c  is, for example, 2.0 mm. One end of the long hole  13   c  is connected to the through hole  13   b  at a position different from the position where one end of the long hole  13   a  is connected to the through hole  13   b . The other end of the long hole  13   c  is formed at the position which is overlapped with the through hole  11   d  in the PDMS block  11 . 
     The through hole  13   d  is formed at the position which is overlapped with the through hole  11   d  in the main body part  3 . The diameter of the through hole  13   d  is, for example, 1.5 mm. 
     The through hole  13   e  is formed at the position which is overlapped with a liver tissue culture chamber  29  in the main body part  3 . The diameter of the through hole  13   e  is, for example, 5.0 mm. 
     The silicone rubber sheet  19  has through holes  19   a ,  19   b ,  19   c  and  19   d , which are, for example, formed by a method such as a punching method. 
     The through hole  19   a  is formed at the position which is overlapped with the end part of the long hole  13   a  in the silicone rubber sheet  13 . The diameter of the through hole  19   a  is, for example, 1.5 mm. 
     The through hole  19   b  is formed at the position which is overlapped with the through hole  13   b  in the silicone rubber sheet  13 . The diameter of the through hole  19   b  is, for example, 4.0 mm. 
     The through hole  19   c  is formed at the position which is overlapped with the end part of the long hole  13   c  in the silicone rubber sheet  13 . The diameter of the through hole  19   c  is, for example, 1.5 mm. 
     The through hole  19   d  is formed at the position which is overlapped with the through hole  13   d  in the silicone rubber sheet  13 . The diameter of the through hole  19   d  is, for example, 1.5 mm. 
     A through hole  19   e  is formed at the position which is overlapped with the through hole  13   e  in the silicone rubber sheet  13 . The diameter of the through hole  19   e  is, for example, 5.0 mm. 
     The filter  15  is, for example, formd of a track-etched membrane PET with a pore size of 1 μm (a product of it4ip s.a.). The diameter of the filter  15  is, for example, 5 mm. The filter  15  is placed between the silicone rubber sheet  13  and the silicone rubber sheet  19  so that the filter  15  is overlapped with the through hole  13   b  and the through hole  19   b.    
     The filter  17  is, for example, formed of a track-etched membrane PC with a pore size of 5 μm (a product of it4ip s.a.). The diameter of the filter  17  is, for example, 6 mm. The filter  17  is placed between the silicone rubber sheet  13  and the silicone rubber sheet  19  so that the filter  17  is overlapped with the through hole  13   e  and the through hole  19   e.    
     For example, after an oxygen plasma treatment is performed on the surfaces of the silicone rubber sheets  13  and  19  on which these sheets are to be stuck together, the silicone rubber sheets  13  and  19  are stuck together in a state where the two filters  15  and  17  are sandwiched by the silicone rubber sheets  13  and  19 . The filter  15  constitutes, for example, a culture surface for an intestinal epithelial cell. The filter  17  constitutes a filter of a culture medium discharging part of the liver tissue culture chamber  29 . 
     The thickness of the PDMS block  21  is, for example, 1.0 mm. The PDMS block  21  has through holes  21   a ,  21   b ,  21   c  and  21   d , as well as a recessed groove  21   e  and a through hole  21   f . For example, the recessed groove  21   e  is formed during the molding of the PDMS block  21 . The through holes  21   a ,  21   b ,  21   c ,  21   d  and  21   f  are formed after the molding of the PDMS block  21 . 
     The through hole  21   a  is formed at the position which is overlapped with the through hole  19   a  in the silicone rubber sheet  19 . The diameter of the through hole  21   a  is, for example, 1.5 mm. 
     The through hole  21   b  is formed at the position which is overlapped with the through hole  19   b  in the silicone rubber sheet  19 . The diameter of the through hole  21   b  is, for example, 4.5 mm. 
     The through hole  21   c  is formed at the position which is overlapped with the through hole  19   c  in the silicone rubber sheet  19 . The diameter of the through hole  21   c  is, for example, 1.5 mm. 
     The through hole  21   d  is formed at the position which is overlapped with the through hole  19   d  in the silicone rubber sheet  19 . The diameter of the through hole  21   d  is, for example, 1.5 mm. 
     The recessed groove  21   e  is formed on the surface to which the silicone rubber sheet  19  is stuck. The depth of the recessed groove  21   e  is, for example, 0.2 mm. The recessed groove  21   e  is formd to be astride the position which is overlapped with both the liver tissue culture chamber  29  and the through hole  19   e  in the silicone rubber sheet  19  and the position which is not overlapped with the liver tissue culture chamber  29 . The depth of the recessed groove  21   e  is, for example, 0.2 mm. A plurality of projections are formed in the position of the recessed groove  21   e  which is overlapped with the through hole  19   e . These projections are provided in order to prevent the filter  17  from sticking to the bottom surface of the recessed groove  21   e.    
     The through hole  21   f  is formed at the position which is overlapped with the recessed groove  21   e  but which is not overlapped with the liver tissue culture chamber  29 . The diameter of the through hole  21   f  is, for example, 1.5 mm. 
     The thickness of the silicone rubber sheet  23  is, for example, 0.2 mm. The silicone rubber sheet  23  has through holes  23   a ,  23   b ,  23   c  and  23   d  as well as a long hole  23   e  and a through hole  23   f.    
     The through hole  23   a  is formed at the position which is overlapped with the through hole  21   a  in the PDMS block  21 . The diameter of the through hole  23   a  is, for example, 1.5 mm. 
     The through hole  23   b  is formed at the position which is overlapped with the through hole  21   b  in the PDMS block  21 . The diameter of the through hole  23   b  is, for example, 4.5 mm. 
     The through hole  23   c  is formed at the position which is overlapped with the through hole  21   c  in the PDMS block  21 . The diameter of the through hole  23   c  is, for example, 1.5 mm. 
     The through hole  23   d  is formed at the position which is overlapped with the through hole  21   d  in the PDMS block  21 . The diameter of the through hole  23   d  is, for example, 1.5 mm. 
     The width dimension of the long hole  23   e  is, for example, 2.0 mm. One end of the long hole  23   e  is connected to the through hole  23   b.    
     The through hole  23   f  is formed at the position which is overlapped with the through hole  21   f  in the PDMS block  21 . The diameter of the through hole  23   f  is, for example, 1.5 mm. 
     The thickness of the PDMS block  25  is, for example, 1.0 mm. The PDMS block  25  has through holes  25   a ,  25   b ,  25   c ,  25   d ,  25   e  and  25   f  as well as recessed grooves  25   g  and  25   h . For example, the recessed grooves  25   g  and  25   h  are formed during the molding of the PDMS block  25 . The through holes  25   a ,  25   b ,  25   c ,  25   d ,  25   e  and  25   f  are formed after the molding of the PDMS block  25 . 
     The through hole  25   a  is formed at the position which is overlapped with the through hole  23   a  in the silicone rubber sheet  23 . The diameter of the through hole  25   a  is, for example, 1.5 mm. 
     The through hole  25   b  is formed at the position which is overlapped with the through hole  23   b  in the silicone rubber sheet  23 . The diameter of the through hole  25   b  is, for example, 4.5 mm. 
     The through hole  25   c  is formed at the position which is overlapped with the through hole  23   c  in the silicone rubber sheet  23 . The diameter of the through hole  25   c  is, for example, 1.5 mm. 
     The through hole  25   d  is formed at the position which is overlapped with the through hole  23   d  in the silicone rubber sheet  23 . The diameter of the through hole  25   d  is, for example, 1.5 mm. 
     The through hole  25   e  is formed at the position which is overlapped with the through hole  23   e  in the silicone rubber sheet  23 . The diameter of the through hole  25   e  is, for example, 1.5 mm. 
     The through hole  25   f  is formed at the position which is overlapped with the through hole  23   f  in the silicone rubber sheet  23 . The diameter of the through hole  25   f  is, for example, 1.5 mm. 
     The recessed grooves  25   g  and  25   h  are formd on the surface which is opposed to the surface to which the silicone rubber sheet  23  is stuck (formed on the surface to which the PDMS block  27  is stuck). The depth of the recessed groove  25   g  is, for example, 0.6 mm. The width dimension of the recessed groove  25   g  is, for example, 2.0 mm. The depth of the recessed groove  25   h  is, for example, 0.2 mm. The width dimension of the recessed groove  25   h  is, for example, 1.7 mm. 
     One end of the recessed groove  25   g  is connected to the through hole  25   b.    
     One end of the recessed groove  25   g  is formed at the position which is overlapped with the penetration groove  25   c . The other end of the recessed groove  25   g  is formd at the position which is overlapped with the penetration groove  25   d.    
     The thickness of the PDMS block  27  is, for example, 1.0 mm. The PDMS block  27  has through holes  27   a ,  27   b ,  27   c  and  27   d . For example, the through holes  27   a ,  27   b ,  27   c  and  27   d  are formed after the molding of the PDMS block  27 . 
     The through hole  27   a  is formed at the position which is overlapped with the through hole  25   a  in the PDMS block  25 . The through hole  27   b  is formed at the position which is overlapped with the through hole  25   f  in the PDMS block  25 . The through hole  27   c  is formd at the position which is overlapped with the tip part of the recessed groove  25   g  in the PDMS block  25 . The through hole  27   d  is formed at the position which is overlapped with the through hole  25   e  in the PDMS block  25 . 
     The diameter of the through holes  27   a ,  27   b ,  27   c  and  27   d  is, for example, 1.5 mm. 
     The forming process of the lid part  5  is briefly explained. 
     The PDMS block  25 , in which the recessed grooves  25   g  and  25   h  are formed, is prepared. In addition, the silicone rubber sheet  23  in which the long hole  23   e  is formed is prepared. The silicone rubber sheet  23  is stuck to the surface of the PDMS block  25  which is opposed to the surface on which the recessed grooves  25   g  and  25   h  are formd. At this time, it is preferable that the surface of the PDMS block  25  to which the silicone rubber sheet  23  is stuck is subjected to an oxygen plasma treatment before being stuck. 
     The through hole  25   e  having a diameter of 1.5 mm is formed to the PDMS block  25  and the silicone rubber sheet  23  after the PDMS block  25  and the silicone rubber sheet  23  are stuck together. 
     The PDMS block  21  in which the recessed groove  21   e  is formed is prepared. The surface of the PDMS block  21  which is opposed to the surface on which the recessed groove  21   e  is formd, and the surface of the silicone rubber sheet  23  which is opposed to the PDMS block  25  are stuck together. At this time, it is preferable that the surface of the PDMS block  21  to which the silicone rubber sheet  23  is stuck is subjected to an oxygen plasma treatment before being stuck. 
     The through holes  21   a ,  23   a , and  25   a , the through holes  21   b ,  23   b , and  25   b , the through holes  21   c ,  23   c , and  25   c , the through holes  21   d ,  23   d , and  25   d  as well as the through holes  21   f ,  23   f , and  25   f  are formed to the PDMS block  21 , the silicone rubber sheet  23  and the PDMS block  25  after the PDMS block  21 , the silicone rubber sheet  23 , and the PDMS block  25  are stuck together. At this time, the diameter of the through holes  21   b ,  23   b , and  25   b  is expanded to 5 mm. The diameter of the through holes  21   a ,  23   a  and  25   a , the through holes  21   c ,  23   c , and  25   c , the through holes  21   d ,  23   d , and  25   d  as well as the through holes  21   f ,  23   f , and  25   f  is 1.5 mm. 
     The silicone rubber sheets  13  and  19 , between which the two filters  15  and  17  are sandwiched, are prepared. To the PDMS block  21 , the silicone rubber sheet  23  and the PDMS block  25  which are stuck together beforehand, the silicone rubber sheet  19  is stuck to the surface of the PDMS block  21  on which the recessed groove  21   e  is formed. 
     The PDMS block  27 , in which the through holes  27   a ,  27   b ,  27   c  and  27   d  are formed, is prepared. To the silicone rubber sheets  13  and  19 , the PDMS block  21 , the silicone rubber sheet  23  and the PDMS block  25  which are stuck together beforehand, the PDMS block  27  is stuck to the silicone rubber sheet  13 . By this, the lid part  5  is completed. 
     In the cell culturing device  1 , the main body part  3  and the lid part  5  are joined by the self-adsorption property of, for example, the PDMS block  11  and the silicone rubber sheet  13 . By this, the main body part  3  and the lid part  5  are stably fixed to each other, for example, without using a special fixing means such as a screw and an adhesive agent. In addition, the main body part  3  can be easily attached/detached to/from the lid part  5 , as needed. 
     In the cell culturing device  1 , the through holes  9   a  and  11   a  in the main body part  3  constitute the liver tissue culture chamber  29  (a first culture chamber). In addition, the through hole  13   e  in the silicone rubber sheet  13  of the lid part  5  constitutes a portion of the liver tissue culture chamber  29 . The bottom surface of the liver tissue culture chamber  29  is constituted by the base plate  7 . The upper surface of the liver tissue culture chamber  29  is constituted by the filter  17 . 
     The through holes  19   b ,  21   b ,  23   b , and  25   b  in the lid part  5  constitute an intestinal epithelial cell culture chamber (a second culture chamber). The lower surface of the intestinal epithelial cell culture chamber  31  is constituted by the filter  15 . The upper surface of the intestinal epithelial cell culture chamber  31  is constituted by the PDMS block  27 . 
     The through holes  27   a ,  25   a ,  23   a , and  21   a , the long hole  13   a , the through hole  13   b , the long hole  13   c , the through holes  19   c ,  21   c ,  23   c , and  25   c , the recessed groove  25   h , the through holes  25   d ,  23   d ,  21   d ,  19   d , and  13   d  in the lid part  5  constitute a culture medium feeding path  33  (a first introduction flow channel). In addition, the through hole  11   d  and the recessed part lib in the main body part  3  also constitute a portion of the culture medium feeding path  33 . The end part of the through hole  27   a  is a culture medium introducing port  33   a.    
     The through hole  19   e , a recessed groove  21   b  as well as the through holes  21   f ,  25   f , and  27   b  in the lid part  5  constitute a culture medium discharging path  35  (a first discharge flow channel). The end part of the through hole  27   b  is a culture medium discharging port  35   a.    
     The through holes  27   c  and  25   i  as well as the recessed groove  25   g  in the lid part  5  constitute a reagent introducing path  37  (a second introduction flow channel). The end part of the through hole  27   c  is a reagent introducing port  37   a.    
     The through holes  27   d  and  25   e  as well as the long hole  23   e  in the lid part  5  constitute a reagent discharging path  39  (a second discharge flow channel). The end part of the through hole  27   d  is a reagent discharging port  39   a.    
     The depth of the recessed groove  25   g  constituting the reagent introducing path  37  is, for example, 0.6 mm. In addition, the depth of the long hole  23   e  constituting the reagent discharging path  39 , which corresponds to the thickness of the silicone rubber sheet  23  is, for example, 0.2 mm. The reason why the height of the flow channel of the reagent introducing path  37  is larger than the height of the flow channel of the reagent discharging path  39  is to make it easy to introduce cells into the intestinal epithelial cell culture chamber  31 . 
     In addition, in the culture medium feeding path  33 , for example, each of the part of the long hole  13   c , the through holes  19   c ,  21   c ,  23   c , and  25   c  and the part of the through holes  25   d ,  23   d ,  21   d ,  19   d , and  13   d  constitutes a liquid collecting part  41 , respectively. The liquid collecting part  41  is provided between the liver tissue culture chamber  29  and the intestinal epithelial cell culture chamber  31  in the culture medium feeding path  33 . 
     The PDMS block  27  which constitutes the upper wall surface of the liquid collecting part  41  is an elastic member which is capable of being penetrated by a suction implement having a sharp tip, in which the through hole is capable of being closed when the suction implement is withdrawn after the penetration due to the elasticity of the member. A liquid, for example, a culture medium is collected from the liquid collecting part  41 , as needed. 
     Meanwhile, though the liquid collecting parts  41  are provided in two spots in this example, the liquid collecting part  41  may be provided in one spot. Further, it is possible to use a part of the recessed groove  25   h  as a liquid collecting part. 
     In the cell culturing device  1 , the main body part  3  in which a liver tissue is cultured can be attached/detached to/from the lid part  5  in which an intestinal epithelial cell is cultured. 
     A cell derived from an intestine, for example, an intestinal epithelial cell (here, Caco2) is introduced together with a culture medium into the intestinal epithelial cell culture chamber  31  in the lid part  5  from a reagent introducing port  39   a . The intestinal epithelial cells are cultured under such a condition that a tight junction is formed on the filter  15 . For example, the intestinal epithelial cells are cultured only in the lid part  5  in a state where the main body part  3  and the lid part  5  are separated from each other. 
     An example of a method for culturing a liver tissue is explained. A gel is introduced as a scaffold into the liver tissue culture chamber  29 , for example, in a state where the main body part  3  is cooled. This gel is, for example, an EHS-gel. The EHS-gel is a preparation of basement membranes isolated from EHS mouse sarcoma cells, and is rich in laminins, collagen type IV and proteoglycans. The EHS-gel liquefies at a low temperature, and solidifies at a normal temperature. Accordingly, the EHS-gel can be solidified on the bottom surface of the liver tissue culture chamber  29  by pouring the EHS-gel into the liver tissue culture chamber  29  in a state where the cell culturing device  1  is cooled, and then leaving the EHS-gel to stand, for example, in an incubator at 37° C. or at room temperature. 
     An endothelial cell (for example, GH7) is seeded on the gel. A network of the endothelial cells is formed on the gel. A hepatic parenchymal cell is seeded on the network of the endothelial cells. In the liver tissue culture chamber  29 , a co-culture system which contains a cell of the endothelial cell lineage and a cell of the hepatocyte lineage is co-cultured on the gel so that the co-culture system has a tubular structure. Examples of the cell of the endothelial cell lineage include, for example, a sinusoidal endothelial cell, a human umbilical vein (artery) endothelial cell, TD2, GH7, and the like. Examples of the cell of the hepatocyte lineage include, for example, ahepatic parenchymal cell, HepaRG, Huh-7, HepG2, TLR2, Hepa 1-6, a liver progenitor cell, and the like. Meanwhile, a cell which is different from a cell of the endothelial cell lineage and a cell of the hepatocyte lineage may be contained in the co-culture system which contains the cell of the endothelial cell lineage and the cell of the hepatocyte lineage. 
     It is confirmed that the intestinal epithelial cells are sufficiently proliferated in the intestinal epithelial cell culture chamber  31  of the lid part  5  and that the tight junction is formed. The main body part  3  in which the liver tissue is cultured in the liver tissue culture chamber  29  and the lid part  5  in which the intestinal epithelial cells are cultured in the intestinal epithelial cell culture chamber  31  are stuck together. The joining surfaces of the main body part  3  and the lid part  5  are both formed of a silicon rubber-based material. Accordingly, the liquid tightness between the main body part  3  and the lid part  5  can be maintained by using a simple jig which is capable of maintaining an appropriate pressure in a state where the main body part  3  and the lid part  5  are stuck together. 
     Further, the protruding part  11   c  having a convex shape is formd on the surface of the PDMS block  11  in the main body part  3  which is in contact with the lid part  5 . The upper surface of the protruding part  11   c  and the silicone rubber sheet  13  in the lid part  5  are joined together. The plane size of the contact surface between the main body part  3  and the lid part  5  is smaller than the plane size of the cell culturing device  1 . Owing to this structure, the contact between the main body part  3  and the lid part  5  at the outer circumferential part of the cell culturing device  1  which is irrelevant to liquid delivery and culture can be eliminated, and a force distribution to an unnecessary part can be suppressed, and therefore, an effect of preventing liquid leakage is exhibited. 
     The culture medium is fed to the culture medium introducing port  33   a , for example, by a syringe pump and the like. The fed culture medium flows into the liver tissue culture chamber  29  via the culture medium feeding path  33  which passes under the filter  15  of the intestinal epithelial cell culture chamber  31  in which intestinal epithelial cells are cultured. This culture medium is discharged from the culture medium discharging port  35   a  via the filter  17  and the culture medium discharging path  35 . 
     A culture medium or a reagent can be fed into the intestinal epithelial cell culture chamber  31  from the reagent introducing port  37   a  via the reagent introducing path  37 . The culture medium and reagent are discharged from the reagent discharging port  39   a  after passing through the intestinal epithelial cell culture chamber  31  via the reagent discharging path  39 . When a reagent is fed into the intestinal epithelial cell culture chamber  31 , a metabolite is passed through the filter  15  at the surface of culture and is supplied to the culture medium feeding path  33  which is connected to the liver tissue culture chamber  29 , after the reagent is absorbed by intestinal epithelial cells. Accordingly, it is important that the intestinal epithelial cells are densely cultured on the filter  15  while forming a tight junction. 
     The metabolite of the drug which is supplied to the culture medium feeding path  33  reaches a liver tissue which is cultured in the liver tissue culture chamber  29  via the culture medium feeding path  33  and the liquid collecting part  41 . This structure makes it possible to simulate a mechanism by which the drug incorporated from the intestinal wall reaches the liver, and also makes it possible to evaluate pharmacokinetics in a state close to in vivo. 
     As described above, the cell culturing device  1  makes it possible to culture two types of cells and to introduce a metabolite of one type of cells into the other type of cells. 
     In addition, in the cell culturing device  1 , the main body part  3  in which the liver tissue culture chamber  29  is formd can be attached/detached to/from the lid part  5  in which the intestinal epithelial cell culture chamber  31  is formed. Therefore, the main body part  3  and the lid part  5  can be combined together at a good timing after culturing each of a liver tissue and an intestinal epithelial cell under an appropriate condition, respectively, in a state where the main body part  3  and the lid part  5  are separated. Accordingly, the cell culturing device  1  enables efficient measurement. 
       FIG. 3  is a schematic diagram for illustrating an example of a cell culturing system which uses the cell culturing device  1  according to the present example. 
     A cell culturing system  43  is a combination of the cell culturing device  1  and a culture medium feeding part which feeds the culture medium into the cell culturing device  1 . The culture medium feeding part has a culture medium housing part  45 , a culture medium feeding pipe  47 , a culture medium discharging pipe  49 , a waste liquid housing part  51 , a liquid feeding pump  53 , and a control part  55 . 
     One end of the culture medium feeding pipe  47  is inserted into the culture medium housing part  45 . The other end of the culture medium feeding pipe  47  is connected to the culture medium introducing port  33   a  of the cell culturing device  1 . One end of the culture medium discharging pipe  49  is connected to the culture medium discharging port  35   a  of the cell culturing device  1 . The other end of the culture medium discharging pipe  49  is inserted into the waste liquid housing part  51 . The liquid feeding pump  53  is connected to the culture medium feeding pipe  47 . The control part  55  controls the operation of the liquid feeding pump  53 . 
     The culture medium housed in the culture medium housing part  45  is sucked by the liquid feeding pump  53 , and is sent to the cell culturing device  1  from the culture medium introducing port  33   a  through the culture medium feeding pipe  47 . The culture medium supplied to the cell culturing device  1  from the culture medium introducing port  33   a  is introduced into the liver tissue culture chamber  29  from the peripheral surface of the liver tissue culture chamber  29  by passing through the culture medium feeding path  33  (see  FIG. 1 ). 
     In accordance with the introduction of the culture medium into the liver tissue culture chamber  29 , apart of the culture medium within the liver tissue culture chamber  29  is discharged from the liver tissue culture chamber  29  to the outside from the culture medium discharging port  35   a  through the filter  17  and the culture medium discharging path  35 . The culture medium discharged from the culture medium discharging port  35   a  is discharged to the waste liquid housing part  51  through the culture medium discharging pipe  49 . 
     When the culture medium within the liver tissue culture chamber  29  is discharged from the culture medium discharging path  35 , the cell culturing device  1  makes the culture medium pass through the filter  17 , and therefore, it is possible to reduce probability of clogging of the culture medium discharging path  35  due to the gel which is peeled off and the like. 
     The cell culturing system  43  is capable of stabilizing environments surrounding cells, tissues, and the like by feeding the culture medium little by little in a manner similar to in vivo. Accordingly, a stable culture of a cell can be realized. As described above, the cell culturing device  1  and the cell culturing system  43  are advantageous for culturing a cell for a long period of time. 
     In addition, when a reagent is supplied into the intestinal epithelial cell culture chamber  31 , a metabolite is supplied to the culture medium feeding path  33  which is connected to the liver tissue culture chamber  29  by passing through the filter  15  on the culture surface, after the reagent is absorbed by the intestinal epithelial cells. The metabolite which is supplied to the culture medium feeding path  33  is supplied into the liver tissue culture chamber  29 , together with the culture medium. Accordingly, the cell culturing device  1  and the cell culturing system  43  make it possible to conduct a test on a reagent in an environment similar to an environment found in vivo. 
     Further, the cell culturing device  1  makes it possible to collect the metabolite which is supplied from the intestinal epithelial cell culture chamber  31  to the culture medium feeding path  33 , together with the culture medium, at the liquid collecting part  41  (see  FIG. 1 ). This collection of the metabolite is, for example, performed by making a suction implement having a sharp tip penetrate the PDMS block  27  which constitutes the upper wall surface of the liquid collecting part  41  in such a way that the suction implement having the sharp tip is inserted into the liquid collecting part  41 , and sucking the metabolite and the culture medium. When the suction implement is withdrawn from the PDMS block  27 , the through hole formed by the penetration of the suction implement is closed by the elastic force of the PDMS block  27 . Accordingly, it is possible to continue supplying the culture medium and the like to the liver tissue culture chamber  29 , even after the metabolite and the like are collected at the liquid collecting part  41 . 
     In the cell culturing system  43  shown in  FIG. 3 , the liquid feeding pump  53  is provided with the culture medium feeding pipe  47 , which has a function which continuously feeds the culture medium into the liver tissue culture chamber  29 , but the cell culturing system according to the present invention is not limited thereto. 
     As an example, as shown in  FIG. 4 , a cell culturing system  57 , which is an example of the present invention, may be a system in which the culture medium discharging pipe  49  is provided with the liquid feeding pump  53 , which has a function which continuously sucks the culture medium from the inside of the liver tissue culture chamber  29 . Meanwhile, the cell culturing system according to the present invention is not limited to the constitution shown in  FIG. 3  or  FIG. 4 . 
     The present invention is capable of stabilizing environments surrounding cells, tissues, and the like by feeding the culture medium little by little in a manner similar to in vivo. Accordingly, the present invention is capable of realizing a stable culture of cells. 
     Incidentally, a body fluid circulates in vivo, while being dialyzed by the kidneys. In that sense, the above-described cell culturing systems  43  and  57  cannot evaluate an effect such as an accumulation of a trace amount of an expression product. 
     Accordingly, by adding a function of dialysis to a halfway part of the cell culturing system according to the present invention, which gives a system in which a culture medium circulates, it is possible to realize an evaluation system closer to an actual living body. 
     According to the present invention, for example, when conducting a pharmacokinetic study, the liver function evaluation considering an influence of an absorption in an intestine is possible, and therefore, a test closer to that conducted in a living body can be realized. In addition, the cell culturing system according to the present invention is a system which utilizes a fluid control technology, and it is possible, for example, to develop a system which considers effects of a dialysis at the kidneys, and further, to develop an integrated system which also considers influences by other organs. 
       FIG. 5  is a schematic diagram for illustrating a further example of a cell culturing system which utilizes the cell culturing device  1  according to the present example. In  FIG. 5 , to the parts which exhibit similar functions to those shown in  FIG. 3  and  FIG. 4 , the same reference signs are given. 
     A culture medium feeding part of a cell culturing system  59  feeds a culture medium into the liver tissue culture chamber  29  of the cell culturing device  1  in a circulated manner. The culture medium feeding part has the culture medium housing part  45 , the culture medium feeding pipe  47 , the culture medium discharging pipe  49 , the liquid feeding pump  53 , the control part  55 , and a dialysis part  61 . 
     One end of the culture medium discharging pipe  49  is connected to the culture medium discharging port  35   a  of the cell culturing device  1 . The other end of the culture medium discharging pipe  49  is connected to the culture medium housing part  45 . The dialysis part  61  is provided in a halfway part of the culture medium discharging pipe  49 . The dialysis part  61  is, for example, a small-sized dialyzer. The dialysis part  61  has a function which filtrates the circulated culture medium such that unnecessary substances are removed. 
     The cell culturing system  59  sucks up the culture medium from the culture medium housing part  45  in which the culture medium is housed, for example, by the liquid feeding pump  53  such as a syringe pump, and supplies the culture medium to the cell culturing device  1 , and thereafter, puts the culture medium back into the original culture medium housing part  45  via the dialysis part  61 . By this, the cell culturing system  59  is capable of evaluating a reagent and the like under an environment closer to that in an actual living body. 
       FIG. 6  is a schematic diagram for illustrating a further example of a cell culturing system which uses the cell culturing device  1  according to the present example. In  FIG. 6 , to the parts which exhibit similar functions to those shown in  FIG. 5 , the same reference signs are given. 
     A cell culturing system  63 , which is an example of the present invention, is different from the cell culturing system  59  shown in  FIG. 5  in the connecting position of the liquid feeding pump  53 . In the cell culturing system  63 , the liquid feeding pump  53  is connected to the culture medium discharging pipe  49  at a position between the cell culturing device  1  and the dialysis part  61 . Meanwhile, the liquid feeding pump  53  may be connected to the culture medium discharging pipe  49  at a position between the dialysis part  61  and the culture medium housing part  45 . 
       FIG. 7  is a schematic diagram for illustrating a further example of a cell culturing system which uses the cell culturing device  1  according to the present example. In  FIG. 7 , to the parts which exhibit similar functions to those shown in  FIG. 3 , the same reference signs are given. 
     A cell culturing system  65 , which is an example of the present invention, further has a dialysis part  67  compared to the cell culturing system  43  shown in  FIG. 3 . The dialysis part  67  is constituted by a dialysis membrane which is provided between two flow channels of the culture medium feeding pipe  47  and of the culture medium discharging pipe  49 . This dialysis membrane is formed of, for example, a Spectra Series (a product of Spectrum, Inc.). 
     The cell culturing device  1 , which is an example of the present invention, is capable of culturing a cell in the liver tissue culture chamber  29  by supplying a culture medium at a flow rate, for example, of about 40 μL/h (microliter/hour), and therefore, the cell culturing device  1  requires a small amount of the culture medium. Accordingly, the cell culturing system  65 , which is an example of the present invention, is also capable of using a dialysis membrane as the dialysis part  67 . 
     Meanwhile, the cell culturing system  65  may feed a dialysate instead of the culture medium to the cell culturing device  1 . 
     In addition, the liquid feeding pump  53  may be connected to the culture medium discharging pipe  49  at a point between the cell culturing device  1  and the dialysis part  61 , as in a cell culturing system  69 , which is an example of the present invention shown in  FIG. 8 . Further, the liquid feeding pump  53  may be connected to the culture medium discharging pipe  49  at a point between the dialysis part  61  and the waste liquid housing part  51 . Furthermore, the liquid feeding pump  53  may be connected to the culture medium feeding pipe  47  at a point between the culture medium housing part  45  and the dialysis part  61 . 
     Incidentally, the cell culturing device  1  explained above has the liquid collecting part  41 . In contrast, it is not necessary for the cell culturing device according to the present invention to have a liquid collecting part. 
       FIG. 9  is a schematic plan view and a cross-sectional schematic diagram for illustrating another example of a cell culturing device.  FIG. 10  is a plan view which shows the cell culturing device of this example by disassembling the cell culturing device. In  FIG. 9  and  FIG. 10 , to the parts which are the same as those shown in  FIG. 1  and  FIG. 2 , the same reference signs are given. 
     A cell culturing device  71  of this example lacks the liquid collecting part  41  compared to the cell culturing device  1  which is explained by referring to  FIG. 1  and  FIG. 2 . That is, the cell culturing device  71  lacks the through holes  19   c ,  21   c ,  23   c  and  25   c  as well as the through holes  13   d ,  19   d ,  21   d ,  23   d  and  25   d  (see  FIG. 1  and  FIG. 2 ). In addition, the cell culturing device  71  lacks the recessed groove  25   h  (see  FIG. 1  and  FIG. 2 ). 
     In the silicone rubber sheet  13  of the cell culturing device  71 , the distal end part of the long hole  13   c  which is opposite to the through hole  13   b  is formed at the position which is overlapped with the through hole lid in the PDMS block  11 . 
     The culture medium feeding path  33  is formd by the through holes  27   a ,  25   a ,  23   a ,  21   a  and  19   a , the long hole  13   a , the through hole  13   b , the long hole  13   c , the through hole lid as well as the recessed part lib. 
     The cell culturing device  71  is capable of exhibiting the same actions and effects as those exhibited by the cell culturing device  1 , which is explained by referring to  FIG. 1  and  FIG. 2 , except for the effects of the liquid collecting part  41  (see  FIG. 1  and  FIG. 2 ). 
     In addition, the cell culturing system, which is explained by referring to, for example,  FIG. 3  to  FIG. 8 , is capable of using the cell culturing device  71  instead of the cell culturing device  1 . 
     The examples of the present invention are explained above, but the constitutions, arrangements, numerical values, and the like in the examples are only given as examples, and the present invention is not limited thereto. Various modifications are possible in the scope of the present invention which is described in claims. 
     As an example, in the above-described examples of the cell culturing device according to the present invention, the first culture chamber is the liver tissue culture chamber  29 , and the second culture chamber is the intestinal epithelial cell culture chamber  31 , but the first culture chamber and the second culture chamber are not limited thereto in the cell culturing device according to the present invention. Cells cultured in the first culture chamber and in the second culture chamber may be any of the various types of cells. 
     In addition, in the above-described examples of the cell culturing device, the culture medium discharging path  35  (the first discharge flow channel) is connected to the liver tissue culture chamber  29  (the first culture chamber) via the filter  17  (the second porous membrane). By this, for example, the clogging of the culture medium discharging path  35  due to the influx of a substance in the liver tissue culture chamber  29  such as a cell and a scaffold material (a gel) into the culture medium discharging path  35  can be prevented. However, it is not necessary to arrange the filter  17  (the second porous membrane) in the cell culturing device according to the present invention. 
     Further, the cell culturing devices  1  and  71  of the above-described examples have the main body part  3  and the lid part  5  which is detachably attached to the main body part  3 . Furthermore, the liver tissue culture chamber  29  (the first culture chamber) is formed in the main body part  3 , and has an opening on the surface of the main body part  3  which is in contact with the lid part  5 . By this, for example, a substance such as a cell, a scaffold material and a culture medium can be placed in the liver tissue culture chamber  29  without interposing the culture medium feeding path  33  (the first introduction flow channel) and the culture medium discharging path  35  (the first discharge flow channel), in a state where the main body part  3  and the lid part  5  are separated. However, the cell culturing device according to the present invention may have a structure which does not have the detachable main body part or the lid part. 
     In addition, in the cell culturing devices  1  and  71  of the above-described examples, the material of a contact surface between the main body part  3  and the lid part  5  is formd of a PDMS or a silicone rubber. By this, the lid part  5  can be stably fixed to the main body part  3  due to the self-adsorption property, for example, without using a special fixing means such as a screw and an adhesive agent. Further, the main body part  3  can be easily attached/detached to/from the lid part  5 , as needed. However, the material of a contact surface between the main body part  3  and the lid part  5  is not limited to a PDMS or a silicone rubber, and materials other than the PDMS and the silicone rubber may be employed. 
     Further, the cell culturing devices  1  and  71  of the above-described examples have the protruding part  11   c  which is provided by forming the surface of the main body part  3  in contact with the lid part  5  in a convex shape. By this, the plane size of the contact surface between the main body part  3  and the lid part  5  is smaller than the plane size of the cell culturing devices  1  and  71 . By making the plane size of the contact surface between the main body part  3  and the lid part  5  smaller, a force distribution to an unnecessary part can be suppressed. By this, the adherence between the main body part  3  and the lid part  5  can be improved. Meanwhile, the convex shape may be formed only on the surface of the lid part  5  in contact with the main body part  3 , or the convex shape may be formed on both the surface of the main body part  3  in contact with the lid part  5  and the surface of the lid part  5  in contact with the main body part  3 . Furthermore, in the cell culturing device according to the present invention, both the surface of the main body part  3  in contact with the lid part  5  and the surface of the lid part  5  in contact with the main body part  3  may be flat surfaces. 
     In addition, in the cell culturing devices  1  and  71  of the above-described examples, the intestinal epithelial cell culture chamber  31  (the second culture chamber) is formed in the lid part  5 . By this, cells can be cultured in the first culture chamber and in the second culture chamber, respectively, under conditions and environments different from each other, by separating the main body part in which the first culture chamber is formed and the lid part in which the second culture chamber is formed. However, the intestinal epithelial cell culture chamber  31  (the second culture chamber) may be formd in the main body part  3 , or the intestinal epithelial cell culture chamber  31  (the second culture chamber) may also be formed to be astride the main body part  3  and the lid part  5 , in the cell culturing device according to the present invention. 
     DESCRIPTION OF REFERENCE SIGNS 
     
         
         
           
               1 : Cell culturing device 
               3 : Main body part 
               5 : Lid part 
               15 : Filter (First porous membrane) 
               17 : Filter (Second porous membrane) 
               29 : Liver tissue culture chamber (First culture chamber) 
               31 : Intestinal epithelial cell culture chamber (Second culture chamber) 
               33 : Culture medium feeding path (First introduction flow channel) 
               35 : Culture medium discharging path (First discharge flow channel) 
               37 : Reagent introducing path (Second introduction flow channel) 
               39 : Reagent discharging path (Second discharge flow channel) 
               41 : Liquid collecting part 
               53 : Liquid feeding pump 
               61 : Dialysis part