CULTURE DEVICE, CULTURE METHOD AND CULTURED ORGAN PRODUCED BY THE CULTURE METHOD

Abstract

A culture device and a culture method by which a substance to be cultured can be three-dimensionally cultured, and the substance thus cultured can be removed as an integrated product with multiple tubes; and a cultured organ produced by the culture method. The culture device comprises a sealed container (2) which contains the substance to be cultured (A) and is disassembled after the culture is completed; multiple ducts (3, 4, 5) arranged in the sealed container (2) and have micropore formed on the outer peripheral surface; a culture medium-feeding device (6, 7) for feeding/circulating the culture medium (B) to at least one duct (3 or 5); an excretory device (8) connected to at least one duct (4) for excreting the waste product (C) permeating into the duct (4) through the micropore of the duct (4) from the substance to be cultured to the outside of the sealed container.

Claims

1-10. (canceled)

11. A cultured organ made to be provided in an organism as an organ of the organism and comprising an integrated product comprising a cultured product of a substance to be cultured and multiple ducts each of which has a plurality of micropores formed on an outer peripheral surface; wherein the multiple ducts comprise at least one first duct made to be inosculated with a blood vessel of the organ in the organism and at least one second duct made to be inosculated with an excretory duct of the organ in the organism; wherein each of the first and second ducts is branched into a plurality of small diameter ducts in the cultured organ, each of the small diameter ducts having the micropores formed on the outer peripheral surface; wherein a structure approximated to blood vessels in the organ in the organism in which the cultured organ is provided is three-dimensionally reproduced by the at least one first duct by configuring the following: the number, mean diameter of the micropores, and quality of material of the first duct; and the number, diameter, way of branching, and arrangement of the small diameter ducts of the first duct; and wherein another structure approximated to excretory ducts in the organ in the organism in which the cultured organ is provided is three-dimensionally reproduced by the at least one second duct by configuring the following: the number, mean diameter of the micropores, and quality of material of the second duct; and the number, diameter, way of branching, and arrangement of the small diameter ducts of the second duct.

12. The cultured organ according to claim 11, wherein a plurality of the first ducts is provided to be made to be inosculated with blood vessels or blood and lymphatic vessels of the organ in the organism, and a structure approximated to blood vessels or blood and lymphatic vessels in the organ in the organism in which the cultured organ is provided is three-dimensionally reproduced by the plurality of first ducts by configuring the following: the number, mean diameter of the micropores, and quality of material of the first ducts; and the number, diameter, way of branching, and arrangement of the small diameter ducts of each of the first ducts.

13. The cultured organ according to claim 12, wherein the cultured organ is used as a liver in the organism, wherein the substance to be cultured is liver cells, wherein the plurality of first ducts are respectively made to be inosculated with a hepatic artery, a portal vein, and a hepatic vein of the liver in the organism, and the at least one second duct is made to be inosculated with a bile duct of the liver in the organism, and wherein a structure approximated to the hepatic artery, the portal vein, and the hepatic vein in the liver in the organism in which the cultured organ is provided is three-dimensionally reproduced by the plurality of first ducts, and another structure approximated to bile ducts in the liver in the organism in which the cultured organ is provided is three-dimensionally reproduced by the at least one second duct.

14. The cultured organ according to claim 12, wherein the cultured organ is used as a pancreas in the organism, wherein the plurality of first ducts are respectively made to be inosculated with blood vessels in the pancreas in the organism, and the at least one second duct is made to be inosculated with an excretory duct of the pancreas connecting the pancreas with a gastrointestinal tract in the organism, and wherein a structure approximated to the blood vessels of the pancreas in the organism in which the cultured organ is provided is three-dimensionally reproduced by the plurality of first ducts, and another structure approximated to excretory ducts of the pancreas connecting the pancreas with a gastrointestinal tract in the organism in which the cultured organ is provided is three-dimensionally reproduced by the at least one second duct.

15. The cultured organ according to claim 11, wherein the cultured organ is produced by culturing three-dimensionally the substance to be cultured by using a culture device comprising a sealed container, wherein the sealed container is for containing the substance to be cultured and has a sealable input port for charging the substance to be cultured and multiple ducts which are arranged in the sealed container and each of which has a plurality of micropores formed on an outer peripheral surface, wherein the multiple ducts of the culture device comprise at least one culture medium-feeding duct for feeding or circulating a culture medium and at least one excretory duct for exhausting a waste product, and wherein the first duct of the cultured organ is originated from the culture medium-feeding duct of the culture device and the second duct of the cultured organ is originated from the excretory duct of the culture device, wherein the culture device further comprises: a culture medium-feeding device connected with the culture medium-feeding duct, wherein the culture medium-feeding device comprises a culture medium circulation circuit for feeding or circulating the culture medium to the culture medium-feeding duct, which is connected to both ends of the culture medium-feeding duct, a pump for circulating the culture medium, which is arranged in the culture medium circulation circuit, and a control box for controlling the circulating culture medium, which is arranged in the culture medium circulation circuit; an excretory device connected with the excretory duct for exhausting waste product permeating into the excretory duct from the substance to be cultured through the micropores of the excretory duct to the outside of the sealed container, wherein the excretory device comprises a waste product-containing vessel arranged in the outside of the sealed container, and a sending duct for sending the waste product accumulating within the excretory duct to the waste product-containing vessel; and at least one partition wall which is arranged in the sealed container so as to divide an inner space of the sealed container into multiple cells and is formed with multiple small holes, the multiple ducts being inserted into some of the multiple small holes in the at least one partition wall so as to be held, and remaining multiple small holes in the at least one partition wall allowing both of the substance to be cultured and the culture medium to migrate between respective cells, wherein the culture medium-feeding duct is branched into a plurality of small diameter ducts in the sealed container, each of the small diameter ducts having the micropores formed on the outer peripheral surface, and the small diameter ducts of the culture medium-feeding duct are respectively inserted into the multiple small holes in the at least one partition wall, wherein the excretory duct is branched into a plurality of small diameter ducts in the sealed container, each of the small diameter ducts having the micropores formed on the outer peripheral surface, and the small diameter ducts of the excretory duct are respectively inserted into the multiple small holes in the at least one partition wall, wherein the culture medium-feeding duct is made to reproduce three-dimensionally a structure approximated to blood vessels in an organ in an organism in which the cultured organ is provided, by configuring the following: the number, mean diameter of the micropores, and quality of material of the culture medium-feeding duct; the number, diameter, way of branching, and arrangement of the small diameter ducts of the culture medium-feeding duct; and arrangement of the multiple small holes for the small diameter ducts of the culture medium-feeding duct in the at least one partition wall, wherein the excretory duct is branched into a plurality of small diameter ducts in the sealed container, each of the small diameter ducts having the micropores formed on the outer peripheral surface, and the small diameter ducts of the excretory duct are respectively inserted into the multiple small holes in the at least one partition wall, wherein the excretory duct is made to reproduce three-dimensionally a structure approximated to excretory ducts in the organ in the organism in which the cultured organ is provided, by configuring the following: the number, mean diameter of the micropores, and quality of material of the excretory duct of the culture device; the number, diameter, way of branching, and arrangement of the small diameter ducts of the excretory duct; and arrangement of the multiple small holes for the small diameter ducts of the excretory duct in the at least one partition wall, and wherein the cultured organ is produced by culturing three-dimensionally the substance to be cultured in the sealed container, while the culture medium is fed to the sealed container through the culture medium-feeding duct by the culture medium-feeding device and the waste product of the substance to be cultured is exhausted to the outside of the sealed container through the excretory duct and the excretory device, while holding the substance to be cultured in the sealed container.

16. The cultured organ according to claim 12, wherein the cultured organ is produced by culturing three-dimensionally the substance to be cultured by using a culture device comprising a sealed container, wherein the sealed container is for containing the substance to be cultured and has a sealable input port for charging the substance to be cultured and multiple ducts which are arranged in the sealed container and each of which has a plurality of micropores formed on an outer peripheral surface, wherein the multiple ducts of the culture device comprise a plurality of culture medium-feeding ducts for feeding or circulating a culture medium and at least one excretory duct for exhausting a waste product, and wherein the plurality of first ducts of the cultured organ is originated from the plurality of culture medium-feeding ducts of the culture device and the second duct of the cultured organ is originated from the excretory duct of the culture device; wherein the culture device further comprises: culture medium-feeding devices respectively connected with the culture medium-feeding ducts, wherein each of the culture medium-feeding devices comprises a culture medium circulation circuit for feeding or circulating the culture medium to the corresponding culture medium-feeding duct, which is connected to both ends of the corresponding culture medium-feeding duct, a pump for circulating the culture medium, which is arranged in the culture medium circulation circuit, and a control box for controlling the circulating culture medium, which is arranged in the culture medium circulation circuit, and wherein the culture medium circulating in each of the culture medium-feeding ducts is the same or are different from the culture medium circulating in another of the culture medium-feeding ducts; an excretory device connected with the excretory duct for exhausting waste product permeating into the excretory duct from the substance to be cultured through the micropores of the excretory duct to the outside of the sealed container, wherein the excretory device comprises a waste product-containing vessel arranged in the outside of the sealed container, and a sending duct for sending the waste product accumulating within the excretory duct to the waste product-containing vessel; and at least one partition wall which is arranged in the sealed container so as to divide an inner space of the sealed container into multiple cells and is formed with multiple small holes, the multiple ducts being inserted into some of the multiple small holes in the at least one partition wall so as to be held, and remaining multiple small holes in the at least one partition wall allowing both of the substance to be cultured and the culture medium to migrate between respective cells; wherein each of the culture medium-feeding ducts is branched into a plurality of small diameter ducts in the sealed container, each of the small diameter ducts having the micropores formed on the outer peripheral surface, and the small diameter ducts of each of the culture medium-feeding ducts are respectively inserted into the multiple small holes in the at least one partition wall; wherein the excretory duct is branched into a plurality of small diameter ducts in the sealed container, each of the small diameter ducts having the micropores formed on the outer peripheral surface, and the small diameter ducts of the excretory duct are respectively inserted into the multiple small holes in the at least one partition wall; wherein the plurality of culture medium-feeding ducts is made to reproduce three-dimensionally a structure approximated to blood vessels or blood and lymphatic vessels in an organ in an organism in which the cultured organ is provided, by configuring the following: the number, mean diameter of the micropores, and quality of material of the culture medium-feeding ducts; the number, diameter, way of branching, and arrangement of the small diameter ducts of each of the culture medium-feeding ducts; and arrangement of the multiple small holes for the small diameter ducts of each of the culture medium-feeding ducts in the at least one partition wall; wherein the excretory duct is branched into a plurality of small diameter ducts in the sealed container, each of the small diameter ducts having the micropores formed on the outer peripheral surface, and the small diameter ducts of the excretory duct are respectively inserted into the multiple small holes in the at least one partition wall; and wherein the excretory duct is made to reproduce three-dimensionally a structure approximated to excretory ducts in the organ in the organism in which the cultured organ is provided, by configuring the following: the number, mean diameter of the micropores, and quality of material of the excretory duct of the culture device; the number, diameter, way of branching, and arrangement of the small diameter ducts of the excretory duct; and arrangement of the multiple small holes for the small diameter ducts of the excretory duct in the at least one partition wall; and wherein the cultured organ is produced by culturing three-dimensionally the substance to be cultured in the sealed container, while the culture medium is fed to the sealed container through each of the culture medium-feeding ducts by a respective culture medium-feeding device and the waste product of the substance to be cultured is exhausted to the outside of the sealed container through the excretory duct and the excretory device, while holding the substance to be cultured in the sealed container.

17. The cultured organ according to claim 15, wherein the cultured organ consists of the integrated product which is obtained by three-dimensionally culturing the substance to be cultured in the sealed container of the culture device in a state that the sealed container containing the substance to be cultured is embedded in the organism and the culture medium-feeding device and the excretory device are arranged in the outside of the organism, and which is comprised of the three-dimensionally cultured product of the substance to be cultured and the multiple ducts, and wherein the integrated product is left with the sealed container in the organism as the organ of the organism.

18. The cultured organ according to claim 15, wherein the sealed container of the culture device is made to be disassembled after culture by cutting at an optional place, or is formed with multiple divided bodies so as to be disassembled after culture, wherein the cultured organ consists of the integrated product which is taken from the sealed container as the integrated product comprising the three-dimensionally cultured product of the substance to be cultured and the multiple ducts by disassembling the sealed container after the culture is completed, and wherein the integrated product is implanted into the organism as the organ of the organism.

19. The cultured organ according to claim 16, wherein the cultured organ consists of the integrated product which is obtained by three-dimensionally culturing the substance to be cultured in the sealed container of the culture device in a state that the sealed container containing the substance to be cultured is embedded in the organism and the culture medium-feeding device and the excretory device are arranged in the outside of the organism, and which is comprised of the three-dimensionally cultured product of the substance to be cultured and the multiple ducts, and wherein the integrated product is left with the sealed container in the organism as the organ of the organism.

20. The cultured organ according to claim 16, wherein the sealed container of the culture device is made to be disassembled after culture by cutting at an optional place, or is formed with multiple divided bodies so as to be disassembled after culture, wherein the cultured organ consists of the integrated product which is taken from the sealed container as the integrated product comprising the three-dimensionally cultured product of the substance to be cultured and the multiple ducts by disassembling the sealed container after the culture is completed, and wherein the integrated product is implanted into the organism as the organ of the organism.

21. The cultured organ according to claim 11, wherein at least one of the multiple ducts is at least partially formed by semipermeable membrane, unglazed cylindrical body, or blood vessel prosthesis.

22. The cultured organ according to claim 11, wherein the plurality of micropores of each of the multiple ducts have a mean diameter of 100 to 10,000 Å.

23. The cultured organ according to claim 17, wherein the sealed container is at least partially formed by a clear material.

24. The cultured organ according to claim 17, wherein the sealed container is in a form of a capsule comprising a cylindrical part and a pair of hemispheres respectively connected to both ends of the cylindrical part.

25. The cultured organ according to claim 17, wherein the sealed container is formed by synthetic resin having flexibility, biomaterial having compatibility to the organism, or material able to melt after a predetermined period in the organism.

26. The cultured organ according to claim 19, wherein the sealed container is at least partially formed by a clear material.

27. The cultured organ according to claim 19, wherein the sealed container is in a form of a capsule comprising a cylindrical part and a pair of hemispheres respectively connected to both ends of the cylindrical part.

28. The cultured organ according to claim 19, wherein the sealed container is formed by synthetic resin having flexibility, biomaterial having compatibility to the organism, or material able to melt after a predetermined period in the organism.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0038] FIG. 1 is a partially broken perspective view showing schematically the first embodiment of the culture device of the present invention.

[0039] FIG. 2 is a plan view showing a part of the second embodiment of the culture device of the present invention.

[0040] FIG. 3 is a longitudinal sectional view with respect to the line III-III in FIG. 2.

[0041] FIG. 4 is a longitudinal sectional view with respect to the line IV-IV in FIG. 2.

[0042] FIG. 5 is an enlarged longitudinal sectional view with respect to the line V-V in FIG. 3.

[0043] FIG. 6 is a perspective view showing an embodiment of partition wall used in the second embodiment.

DESCRIPTION OF EMBODIMENTS

[0044] Now, the first embodiment of the culture device of the present invention is explained with referring the FIG. 1.

[0045] The culture device comprises a sealed container 2 having a sealable input port 1 for charging the substance to be cultured A (multiple input ports may be used), said sealed container being able to contain the substance to be cultured A and to be disassembled after the culture; multiple ducts 3, 4, 5 arranged in the sealed container 2, each of ducts having micropore (not shown) formed on the outer peripheral surface; culture medium-feeding devices 6, 7 connected to at least one of the ducts 3, 4, 5, in this embodiment, two ducts 3, 5, respectively, the respective culture medium-feeding devices feeding or circulating the culture medium B to the duct 3, 5, so that the ducts form two culture medium-feeding ducts; and an excretory device 8 connected to at least one duct 4 other than the culture medium-feeding ducts 3, 5 among the ducts 3, 4, 5, said excretory device exhausting the waste product permeating into the duct 4 from the substance to be cultured A through the micropore of the duct to the outside of the sealed container 2, so that the duct 4 forms an excretory duct.

[0046] The input port 1 of the sealed container 2 can be sealed by a sealable lid 1a after charging the substance to be cultured A to the sealed container 2. Alternatively, the sealable lid 1a may be made of an elastic material such as rubber, etc. and the substance to be cultured A such as cell, etc. may be charged into the sealed container 2 via an injection needle, etc. (not shown) stuck into the sealable lid.

[0047] The sealed container 2 is a rectangular parallelepiped box made of clear acrylic plate, or glass plate, etc. The sealed container may be in the form of vertical or horizontal type cylinder, or other shapes.

[0048] It is possible to form only a part of the sealed container 2 by a clear material and to form the remaining by an opaque material.

[0049] According to this embodiment, the sealed container 2 is-made of acrylic plate and after the culture it can be disassembled by cutting it at an optional place with a cutter, etc.

[0050] The sealed container 2 may be formed by using multiple divided bodies (not shown) and by inserting a packing into between the divided bodies and connecting them with bolt and nut. After the culture, the divided bodies may be disassembled by unlocking the bolt and nut or removing the connecting portion.

[0051] The conditions of duct 3, 4, 5 such as number, length, outer diameter, inner diameter, quality of material, diameter of fine pore, etc. are determined in accordance with the culture condition.

[0052] For example, the duct 3, 4, 5 is selected from cylindrical body made of semi-permeable membrane or unglazed earthenware, blood vessel prosthesis, etc. The duct 3, 4, 5 may be formed by the cylindrical body made of semi-permeable membrane or unglazed earthenware only at its portion present within the sealed container 2 and by a cylindrical body made of synthetic resin at its other portion.

[0053] The duct 3, 4, 5 has preferably the outer diameter of several mm to tens mm and the mean diameter of fine pore of 100 to 10,000 Å.

[0054] The ducts 3, 4, 5 may differ from each other in quality of material, size, and mean diameter of fine pore, etc. in accordance with a use of duct.

[0055] It is desirable that the duct 3, 4, 5 has flexibility, but that is not essential.

[0056] According to this embodiment, the respective ducts 3, 4, 5 penetrate in horizontal direction through the sealed container 2 and their both ends project from both sides of the sealed container 2.

[0057] In order to prevent the culture medium B, etc. mentioned below from leaking from the respective ducts 3, 4, 5 situated at the outside of the sealed container, the area wherein the micropore is formed in the respective ducts 3, 4, 5 is limited, or the micropore is formed throughout the respective ducts 3, 4, 5, following by coating airtightly or liquid-tightly the periphery of the respective ducts situated at the outside of the sealed container 2 or by fitting externally a flexible tube (not shown) to the respective ducts situated in the outside of the sealed container 2.

[0058] The respective culture medium-feeding devices 6, 7 comprise a culture medium circulation circuit 9 for circulating the culture medium B to the culture medium-feeding duct 3, 5, said culture medium circulation circuit 9 being connected to both ends of the culture medium-feeding duct 3, 5; a pump 10 for circulating the culture medium B, said pump 10 being arranged in the culture medium circulation circuit 9; and a control box 11 for controlling the temperature, pressure, oxygen concentration, ingredient distribution, etc. of the circulating culture medium B, said control box being arranged in the culture medium-feeding circulation circuit 9.

[0059] The respective culture medium-feeding devices 6, 7 comprise a liquid reservoir 12 in the culture medium circulation circuit 9, the culture medium circulation circuit 9 comprising a liquid-feeding duct 9a leading from the liquid reservoir 12 to one end of the respective ducts 3, 5 via the control box 11 and the pump 10; and a liquid-exhausting duct 9b leading from the other end of the respective ducts 3, 5 to the liquid reservoir 12.

[0060] An excretory device 8 comprises a waste product-containing vessel 13 arranged in the outside of the sealed container 2; and a sending duct 14 for sending the waste product C accumulating within the excretory duct 4 to the waste product-containing vessel 13.

[0061] The other end of the excretory duct 4 is closed by a plug 15.

[0062] Now, an embodiment of the culture method of the present invention which is carried out by using the above-mentioned culture device is explained.

[0063] As being shown in FIG. 1, the input port 1 of the sealed container 2 is open and the substance to be cultured A is charged into the sealed container 2 and then the input port 1 is sealed by the sealable lid 1a.

[0064] The substance to be cultured A may be as follows:

[0065] (1) multiple cells dispersed in culture medium of liquid type or of gel type and floating therein;

[0066] (2) small aggregate of cells floating in culture medium of liquid type or of gel type.

[0067] After the substance to be cultured A is charged into the sealed container 2 and the input port 1 is sealed, the culture medium B is circulated into the culture medium-feeding ducts 3, 5 by the culture medium-feeding devices 6, 7.

[0068] Then, the essential ingredients contained in the culture medium B permeate to the side of the substance to be cultured A within the sealed container 2 through the micropore of the respective culture medium-feeding ducts 3, 5 so that the culture of the substance to be cultured A is prompted.

[0069] The culture medium B is selected from well-known preparations containing one or two or more of collagen, elastin, proteoglycan, fibrillin, fibronectin, laminin, chitin, chitosan, blood, etc.

[0070] The waste product C of the substance to be cultured A permeates into the excretory duct 4 through the micropore of the excretory duct 4 and is sent to the waste product-containing vessel 13 through the feeding duct 14 and is stored in the waste product-containing vessel 13.

[0071] Thus, the waste product C of the substance to be cultured A is exhausted to the outside of the sealed container 2 through the excretory duct 4 and feeding duct 14 and accordingly, the waste product C do not stay in the substance to be cultured A, whereby it is possible to carry out effectively the three-dimensional culture for a prolonged period.

[0072] According to the culture method, it is possible to culture three-dimensionally the substance to be cultured A for a prolonged period in the sealed container 2 under the conditions approximate to the complicated structure of various organs in the organism of animal.

[0073] It is possible to take out the cultured product of the substance to be cultured A and the ducts 3, 4, 5 as an integrated product by disassembling the sealed container 2 after the culture is completed. The resultant integrated product can be implanted into the organism as cultured organ just as it is.

[0074] At that time, the culture medium-feeding ducts 3, 5 can serve as blood vessel. Accordingly, the integrated product after the implantation can maintain the function similar to the various organs of animal such as liver, pancreas, etc. by inosculating the culture medium-feeding ducts 3, 5 to blood vessel and inosculating the excretory duct 4 to gastrointestinal tract, bile duct, or the like.

[0075] FIGS. 2 to 6 show the second embodiment of the culture device of the present invention.

[0076] In the second embodiment, the sealed container 20 is in the form of capsule consisting of hemispheres 20b, 20b connected to the both ends of cylindrical part 20a, said cylindrical part being provided at an optional position with the input port 21 for charging the substance to be cultured A into the sealed container 20.

[0077] The sealable lid 22 is arranged detachably to the input port 21.

[0078] It is preferable that when the sealable lid 22 is mounted suitably on the input port 21, the surface of the sealable lid 22 matches with the outer surface of the cylindrical part 20a of the sealed container 20.

[0079] The sealed container 20 and sealable lid 22 are preferably formed by synthetic resin having flexibility, biomaterial having compatibility to the organism such as collagen fiber, etc., or material able to melt after the period longer than the predetermined culture period lapse on being embedded in the organism. There are exemplified as actual material nylon, reinforced glass, reinforced plastics, silicone rubber, silicone prosthesis, etc.

[0080] As being shown in FIG. 3 and FIG. 6, multiple partition walls 23 made of the same biomaterial as above-mentioned biomaterial are arranged within the sealed container 20 at the suitable intervals in the horizontal direction.

[0081] The respective partition walls 23 have multiple small holes 24 arranged in the predetermined configuration or random configuration.

[0082] The inner space of the sealed container 20 is divided into multiple cells 25 by the respective partition walls 23.

[0083] The multiple small holes 24 formed in the respective partition walls 23 allow the substance to be cultured A and the culture medium B to migrate between respective cells 25, 25 and have the function of holding the below-mentioned small diameter ducts 23a-26a inserted into any of small holes 24 at the predetermined position in the sealed container 20. The shape and size of the multiple small holes 24 formed in the respective partition walls 23 are suitably modifiable. It is possible to block the migration of substance to be cultured A and culture medium B between the respective cells 25, 25 by the partition walls 23.

[0084] The sealed container 20 has four ducts 26-29 arranged in the manner that they penetrate through the sealed container in longitudinal direction.

[0085] In the second embodiment, as being shown in FIG. 3, the respective ducts 26-29 consist of multiple large diameter ducts 26a-29a penetrating through the hemisphere parts 20b at the both ends of the sealed container 20, while maintaining airtightness and liquid-tightness, and multiple small diameter ducts 26b-29b which are mutually bundled at their both ends in the sealed container 20 and are, as being shown in FIG. 3 and FIG. 5, connected to the large diameter ducts 26a-29a by a connecting pipe 30 made of elastic material such as rubber, etc.

[0086] The peripheral surface of respective small diameter ducts 26b-29b is provided with the micropore (not shown) similar to the micropore formed in the ducts 3, 4, 5 of the first embodiment.

[0087] As being shown in FIG. 5, it is preferable to keep the airtightness and liquid-tightness in the connecting part by providing the sealing material 31 into the space formed by the bundling part in which the ends of multiple small diameter ducts 26b-29b are mutually bundled and the space formed by the connecting pipe 30 and the bundling part.

[0088] The respective small diameter ducts 26b-29b are held at the predetermined position by penetrating through any of the small holes 24 of the respective partition walls 23, so that they do not get entangled.

[0089] Omitting to show in the figure, both ends of three ducts among the four ducts 26-29, for example, ducts 26, 28, 29 are connected to a culture medium-feeding device (not shown) similar to the culture medium-feeding device 6 or 7 connected to the duct 3 or 5 in the first embodiment. One end of remaining duct 27 is connected to an excretory device (not shown) similar to the excretory device 8 connected to the duct 4 in the first embodiment and another end of the duct 27 is closed by a plug (not shown) pressed into the end, which is similar to the plug 15 in the first embodiment.

[0090] Accordingly, in this embodiment, the ducts 26, 28 and 29 serve as a culture medium-feeding duct and the duct 27 serves as an excretory duct.

[0091] In order to facilitate the engraftment of cell within the sealed container 20 and to prevent the duct 26, 27, 28, 29 from clogging by blood cake, etc., it is desirable to enclose fiber for engraftment in the sealed container 20, or to implant fine fiber on inner surface of the sealed container 20 or surface of the partition wall 23, etc., or to cover the surfaces with a material capable of easy engraftment of cell or to cover the inner surface of the duct 26, 27, 28, 29 with a material which renders engraftment of cell difficult.

[0092] Now, an embodiment of the culture method of the present invention, which is carried out by using the second embodiment of the above-mentioned culture device is explained.

[0093] In the state shown in FIGS. 2-4, the sealable lid 22 is removed from the input port 21 of the sealed container 20 to open the input port 21. The substance to be cultured A is charged into the sealed container 20 therefrom and the sealable lid 22 is mounted on the input port 21 to seal the input port 21.

[0094] Then, both ends of the respective ducts 26, 28, 29 are connected to the culture medium-feeding device, and one end of the duct 27 is connected to the excretory device and the plug is compressed into another end of the duct 27 to close the end.

[0095] The sealed container 20 and the ducts 26-29 connected to the sealed container are embedded in the organism of animal (not shown) as it is in this state.

[0096] Then, the pump in the respective culture medium-feeding devices (refer to the pump 10 in FIG. 1) is actuated to circulate the culture medium B similar to the above-mentioned culture medium to the ducts 26, 28, 29.

[0097] The culture medium B circulating in the respective ducts 26, 28, 29 is same as each other or may be different from each other.

[0098] In this way, the waste product C of the substance to be cultured A, which permeates into the small diameter duct 27b through micropore of the small diameter duct 27b of the excretory duct 27, is exhausted to the outside of the sealed container 20 via the large diameter duct 27a, while circulating the culture medium B to the respective culture medium-feeding ducts 26, 28, 29. As a result, as with the first embodiment, it is possible to culture three-dimensionally and effectively the substance to be cultured A in the sealed container 20 for a prolonged period, while the waste product C do not stay in the substance to be cultured A.

[0099] According to this culture method, the sealed container 20 is embedded in the organism and the three-dimensional culture of the substance to be cultured A is conducted therein and accordingly, it is possible to maintain the same conditions such as temperature, etc. as the organism.

[0100] After the culture is completed, when, for example, the ducts 26, 28, 29, and 27 are inosculated to the blood vessel, portal vein, bile duct, and lever hepatic vain (in the case that the substance to be cultured A is liver cell) of the organism, respectively and the sealed container 20 is left in the organism, it is possible to use the cultured cell as a part of the organism just as it is.

[0101] In this case, when the sealed container 20 is made of a material capable of melting in the organism after passing definite period of time, it is not required to take out the sealed container 20 from the organism.

[0102] The number of the ducts 26-29, number and diameter of the small diameter ducts 26b-29b with respect to the large diameter ducts 26a-29a in the respective ducts 26-29, how to branch, arrangement thereof, etc. may be varied in accordance with type of the substance to be cultured A, condition of recipient, etc.

[0103] The present invention is not limited only to the above-mentioned embodiments, and can be carried out in accordance with the following many modified embodiments without departing the scope and spirit of the Claims.

[0104] (1) One end of the culture medium-feeding duct 3 or 5 is closed and the culture medium B is fed to the sealed container via another end.

[0105] (2) Either of the ducts 3 and 5 is omitted and accordingly either of the culture medium-feeding devices 6 and 7 is omitted.

[0106] (3) Either of the ducts 3 and 5 is used as lymphatic vessel.

[0107] (4) The ducts 3, 4 and 5 are used as blood duct, excretory duct, and lymphatic vessel, respectively.

[0108] (5) The substance to be cultured A is three-dimensionally cultured, while reproducing the complicated structure similar to the structure of organs in the organism in the points of complicated extending of blood vessels, flow of blood and bile, by using one duct as the excretory duct 4 and multiple ducts as the culture medium-feeding duct 3, 5 and by modifying the layout of ducts.

[0109] (6) The respective ducts 3, 4, 5 are arranged in the different direction in the sealed container 20.

REFERENCE SIGNS LIST

[0110] A substance to be cultured [0111] B culture medium [0112] C waste product [0113] 1 input port [0114] 1a salable lid [0115] 2 sealed container [0116] 3 duct (culture medium-feeding duct) [0117] 4 duct (excretory duct) [0118] 5 duct (culture medium-feeding duct) [0119] 6, 7 culture medium-feeding device [0120] 8 excretory device [0121] 9 culture medium circulation circuit [0122] 9a liquid-feeding duct [0123] 9b liquid-excreting duct [0124] 10 pump [0125] 11 control box [0126] 12 liquid reservoir [0127] 13 waste product containing vessel [0128] 14 feeding duct [0129] 15 plug [0130] 20 sealed container [0131] 20a cylindrical part [0132] 20b hemisphere part [0133] 21 input port [0134] 22 salable lid [0135] 23 partition wall [0136] 24 small pore [0137] 25 cell [0138] 26, 28, 29 duct (culture medium-feeding duct) [0139] 27 duct (excretory duct) [0140] 26a-29a large diameter duct [0141] 26b-29b small diameter duct [0142] 30 connecting duct [0143] 31 sealing material