Creation of tissue engineered female reproductive organs

• US 20030096407A1
• Filed: 11/15/2002
• Published: 05/22/2003
• Est. Priority Date: 11/16/2001
• Status: Active Grant

• Current Assignee: Anthony J. Atala
• Original Assignee: Children's Medical Center Corporation
• Inventors: Atala, Anthony, Yoo, James J.
• Granted Patent: US 7,806,937 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61L 2430/22 for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3679 Hollow organs, e.g. bladder, esophagus, urether, uterus, intestine
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/3813 Epithelial cells, e.g. keratinocytes, urothelial cells
• CPC Class Codes: A61L 27/3826 Muscle cells, e.g. smooth muscle cells
• CPC Class Codes: A61L 27/3886 comprising two or more cell types
• CPC Class Codes: A61L 27/3891 as distinct cell layers
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2502/1347 Smooth muscle cells
• CPC Class Codes: C12N 2502/243 Cells of the female genital tract, non-germinal ovarian cells
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691

Abstract

The invention is directed to compositions and methods for reconstructing artificial female reproductive organs. The constructs and methods of the invention can be used for ameliorating congenital malformations and disorders of female reproductive tract using tissue engineered female reproductive organs, such as the uterus, vagina, cervix, and fallopian tubes. These tissue engineered female reproductive organs can be generated by perfusing cultured cell populations derived from cells of the female reproductive tissues, such as uterine, vaginal, cervical, fallopian tube epithelial cells as well as smooth muscle cells.

Claims

1. A method of reconstructing a female reproductive organ or tissue structures comprising:

providing a biocompatible matrix;

perfusing the biocompatible matrix with a first cell population, the first cell population comprising a female reproductive cell population;

perfusing the biocompatible matrix with a second cell population comprising a different cell type than the first cell population; and

culturing the cell populations in the biocompatible matrix;

to thereby create a female reproductive organ, tissue structure, or segment thereof.

2. The method of claim 1, wherein the method further comprises perfusing the biocompatible matrix with at least one additional cell population.

3. The method of claim 1, wherein the method further comprises implanting the cell-perfused biocompatible matrix into a subject for the formation of the female reproductive organ or tissue structure.

4. The method of claim 1, wherein the female reproductive organ is selected from the group consisting of uterus, vagina, fallopian tube, and cervix.

5. The method of claim 1 wherein the first cell population comprises a substantially homogenous female reproductive epithelial cell population.

6. The method of claim 1 wherein the first cell population comprises a substantially homogenous vaginal epithelial cell population.

7. The method of claim 1 wherein the second cell population comprises a substantially homogenous smooth muscle cell population.

8. The method of claim 1, wherein the first and second cell populations are deposited on separate biocompatible matrix layers and the biocompatible matrix layers are combined after the perfusion steps.

9. The method of claim 1, wherein the female reproductive organ or tissue has epithelial cells deposited on a surface of the matrix and smooth muscle cells deposited on another surface of the matrix.

10. The method of claim 1, wherein the biocompatible matrix comprises a polymer.

11. The method of claim 1, wherein the biocompatible matrix comprises a decellularized structure.

12. The method of claim 1, wherein the biocompatible matrix is formed from at least one material selected from the group consisting of cellulose ether, cellulose, cellulosic ester, fluorinated polyethylene, poly-4-methylpentene, polyacrylonitrile, polyamide, polyamideimide, polyacrylate, polybenzoxazole, polycarbonate, polycyanoarylether, polyester, polyestercarbonate, polyether, polyetheretherketone, polyetherimide, polyetherketone, polyethersulfone, polyethylene, polyfluoroolefin, polyglycolic acid, polyimide, polyolefin, polyoxadiazole, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfide, polysulfone, polytetrafluoroethylene, polythioether, polytriazole, polyurethane, polyvinyl, polyvinylidene fluoride, regenerated cellulose, silicone, urea-formaldehyde, copolymers and physical blends thereof.

13. The method of claim 1 wherein the biocompatible matrix is polyglycolic acid (PGA).

14. The method of claim 1 wherein the biocompatible matrix is coated with a biocompatible and biodegradable shaped setting material.

15. A method of reconstructing, repairing, augmenting or replacing a female reproductive organ or tissue structures comprising:

providing a biocompatible matrix;

perfusing the biocompatible matrix with a first cell population, the first cell population comprising a female reproductive cell population;

perfusing the biocompatible matrix with a second cell population comprising a different cell type than the first cell population;

culturing the cell populations in the biocompatible matrix; and

implanting the biocompatible matrix into a subject;

to thereby create a female reproductive organ, tissue structure, or segment thereof in the subject in need of reconstruction, repair, augmentation or replacement of the female reproductive organ or tissue structure.

16. The method of claim 15, wherein the method further comprises perfusing the biocompatible matrix with at least one additional cell population.

17. The method of claim 15, wherein the female reproductive organ is selected from the group consisting of uterus, vagina, fallopian tube, and cervix.

18. The method of claim 15, wherein the first cell population comprises a substantially homogenous female reproductive epithelial cell population.

19. The method of claim 15, wherein the first cell population comprises a substantially homogenous vaginal epithelial cell population.

20. The method of claim 15, wherein the second cell population comprises a substantially homogenous smooth muscle cell population.

21. The method of claim 15, wherein the first and second cell populations are perfused on separate matrix layers and the matrix layers are combined after perfusion.

22. The method of claim 15, wherein the biocompatible matrix comprises a polymer.

23. The method of claim 15, wherein the biocompatible matrix comprises a decellularized structure.

24. The method of claim 15, wherein the biocompatible matrix is formed from at least one material selected from the group consisting of cellulose ether, cellulose, cellulosic ester, fluorinated polyethylene, poly-4-methylpentene, polyacrylonitrile, polyamide, polyamideimide, polyacrylate, polybenzoxazole, polycarbonate, polycyanoarylether, polyester, polyestercarbonate, polyether, polyetheretherketone, polyetherimide, polyetherketone, polyethersulfone, polyethylene, polyfluoroolefin, polyglycolic acid, polyimide, polyolefin, polyoxadiazole, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfide, polysulfone, polytetrafluoroethylene, polythioether, polytriazole, polyurethane, polyvinyl, polyvinylidene fluoride, regenerated cellulose, silicone, urea-formaldehyde, copolymers and physical blends thereof.

25. The method of claim 15, wherein the biocompatible matrix is polyglycolic acid (PGA).

26. A method of constructing an artificial vagina comprising:

providing a biocompatible matrix;

perfusing the biocompatible matrix with a first cell population, the first cell population comprising a vaginal epithelial cell population;

perfusing the biocompatible matrix with a second cell population of a different cell type than the first cell population; and

culturing the cell populations in the biocompatible matrix;

to thereby create an artificial vagina, vaginal tissue structure, or segment thereof.

27. The method of claim 26, wherein the method further comprises perfusing the biocompatible matrix with at least one additional cell population.

28. The method of claim 26, wherein the second cell population comprises a substantially homogenous smooth muscle cell population.

29. The method of claim 26, wherein the biocompatible matrix comprises a polymer.

30. The method of claim 26, wherein the biocompatible matrix comprises a decellularized structure.

31. The method of claim 26, wherein the biocompatible matrix is formed from at least one material selected from the group consisting of cellulose ether, cellulose, cellulosic ester, fluorinated polyethylene, poly-4-methylpentene, polyacrylonitrile, polyamide, polyamideimide, polyacrylate, polybenzoxazole, polycarbonate, polycyanoarylether, polyester, polyestercarbonate, polyether, polyetheretherketone, polyetherimide, polyetherketone, polyethersulfone, polyethylene, polyfluoroolefin, polyglycolic acid, polyimide, polyolefin, polyoxadiazole, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfide, polysulfone, polytetrafluoroethylene, polythioether, polytriazole, polyurethane, polyvinyl, polyvinylidene fluoride, regenerated cellulose, silicone, urea-formaldehyde, copolymers and physical blends thereof.

32. The method of claim 26, wherein the biocompatible matrix is polyglycolic acid (PGA).

33. A method of reconstructing, repairing, augmenting or replacing a vagina in a subject comprising:

providing a biocompatible matrix;

perfusing the biocompatible matrix with a first cell population, the first cell population comprising a vaginal epithelial cell population;

perfusing the biocompatible matrix with a second cell population comprising a different cell type than the first cell population;

culturing the cell populations in the biocompatible matrix; and

implanting the cell perfused biocompatible matrix in the subject;

to thereby create an artificial vagina or vaginal tissue structure in the subject in need of vaginal reconstruction, repair, augmentation or replacement.

34. The method of claim 33, wherein the method further comprises perfusing the biocompatible matrix with at least one additional cell population.

35. The method of claim 33, wherein the second cell population comprises a substantially homogenous smooth muscle cell population.

36. The method of claim 33, wherein the biocompatible matrix comprises a decellularized structure.

37. The method of claim 33, wherein the biocompatible matrix is formed from at least one material selected from the group consisting of cellulose ether, cellulose, cellulosic ester, fluorinated polyethylene, poly-4-methylpentene, polyacrylonitrile, polyamide, polyamideimide, polyacrylate, polybenzoxazole, polycarbonate, polycyanoarylether, polyester, polyestercarbonate, polyether, polyetheretherketone, polyetherimide, polyetherketone, polyethersulfone, polyethylene, polyfluoroolefin, polyglycolic acid, polyimide, polyolefin, polyoxadiazole, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfide, polysulfone, polytetrafluoroethylene, polythioether, polytriazole, polyurethane, polyvinyl, polyvinylidene fluoride, regenerated cellulose, silicone, urea-formaldehyde, copolymers and physical blends thereof.

38. The method of claim 33, wherein the biocompatible matrix is polyglycolic acid (PGA).

39. A construct for the reconstruction, repair, augmentation or replacement of a female reproductive organ or tissue structure comprising:

an implantable, biocompatible matrix, and at least two different cell populations, which have been deposited on or in the biocompatible matrix and cultured to form the female reproductive organ or tissue;

wherein at least one of the two different cell populations comprises a female reproductive cell population.

40. The construct of claim 39, wherein the biocompatible matrix is formed from at least one material selected from the group of materials consisting of cellulose ether, cellulose, cellulosic ester, fluorinated polyethylene, poly-4-methylpentene, polyacrylonitrile, polyamide, polyamideimide, polyacrylate, polybenzoxazole, polycarbonate, polycyanoarylether, polyester, polyestercarbonate, polyether, polyetheretherketone, polyetherimide, polyetherketone, polyethersulfone, polyethylene, polyfluoroolefin, polyglycolic acid, polyimide, polyolefin, polyoxadiazole, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfide, polysulfone, polytetrafluoroethylene, polythioether, polytriazole, polyurethane, polyvinyl, polyvinylidene fluoride, regenerated cellulose, silicone, urea-formaldehyde, copolymers and physical blends thereof.

41. The construct of claim 39, wherein the biocompatible matrix is polyglycolic acid (PGA).

42. The construct of claim 39, wherein the biocompatible matrix comprises a decellularized structure.

43. The construct of claim 39, wherein the first cell population comprises a substantially homogenous female reproductive epithelial cell population.

44. The construct of claim 39, wherein the first cell population comprises a substantially homogenous vaginal epithelial cell population.

45. The construct of claim 39, wherein the second cell population comprises a substantially homogenous smooth muscle cell population.

46. The construct of claim 39, wherein the female reproductive organ or tissue structure is selected from the group consisting of vagina, uterus, fallopian tube, and cervix.

47. The construct of claim 39, wherein the female reproductive organ or tissue structure is a vagina or vaginal segment and having epithelial cells deposited on a surface of the matrix and smooth muscle cells deposited on another surface of the matrix.

48. The construct of claim 39, wherein the first and second cell populations are deposited sequentially.

49. The construct of claim 39, wherein the first and second cell populations are deposited on separate matrix layers and the matrix layers are combined after the deposition steps.

50. An artificial vagina construct comprising:

an implantable, biocompatible matrix, which has been perfused with a vaginal epithelial cell population and with a smooth muscle cell population, such that the vaginal epithelial cell population attaches to the smooth muscle cell population forming a tissue structure having compliance of normal vaginal tissue.

51. The construct of claim 50, wherein the biocompatible matrix is formed from at least one material selected from the group consisting of cellulose ether, cellulose, cellulosic ester, fluorinated polyethylene, poly-4-methylpentene, polyacrylonitrile, polyamide, polyamideimide, polyacrylate, polybenzoxazole, polycarbonate, polycyanoarylether, polyester, polyestercarbonate, polyether, polyetheretherketone, polyetherimide, polyetherketone, polyethersulfone, polyethylene, polyfluoroolefin, polyglycolic acid, polyimide, polyolefin, polyoxadiazole, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfide, polysulfone, polytetrafluoroethylene, polythioether, polytriazole, polyurethane, polyvinyl, polyvinylidene fluoride, regenerated cellulose, silicone, urea-formaldehyde, copolymers and physical blends thereof.

52. The construct of claim 50, wherein the biocompatible material comprises a decellularized structure.

53. The construct of claim 50, wherein the biocompatible material is polyglycolic acid (PGA).

54. A method of reducing infertility in a subject comprising:

providing a biocompatible matrix;

perfusing the biocompatible matrix with a first cell population, the first cell population comprising a female reproductive cell population;

perfusing the biocompatible matrix with a second cell population comprising a different cell type than the first cell population;

culturing the cell populations in the biocompatible matrix; and

implanting the cell perfused biocompatible matrix in the subject;

to thereby create an artificial female reproductive organ, tissue structure, or segment thereof in the subject, whereby the artificial organ, tissue or segment thereof, reduces infertility in the subject.

55. The method of claim 54, wherein the female reproductive organ is selected from the group consisting of vagina, fallopian tube, cervix and uterus.

56. The method of claim 54, wherein the first cell population comprises a substantially homogenous female reproductive epithelial cell population.

57. The method of claim 54, wherein the first cell population is selected from the group consisting of vaginal epithelial cell population, fallopian tube epithelial cell population, cervical epithelial cell population, and uterine epithelial cell population.

58. The method of claim 54, wherein the second cell population comprises a substantially homogenous smooth muscle cell population.

59. The method of claim 54, wherein the method further comprises forming an artificial female reproductive organ or tissue structure that exhibits the compliance of natural female reproductive organ.

60. The method of claim 54, wherein the method further comprises impregnating the subject with a fertilized egg.

61. A method of screening a compound that modulates female reproductive cells comprising:

providing a biocompatible matrix, which has been perfused with a female reproductive epithelial cell population and with a smooth muscle cell population, such that the female reproductive epithelial cell population attaches to the smooth muscle cell population forming an artificial female reproductive tissue;

contacting the artificial female reproductive tissue with a library of test compounds; and

selecting from the library of test compounds a compound of interest that modulates female reproductive cells.

62. The method of claim 61, wherein the compound is cytotoxic to the female reproductive cells.

63. The method of claim 61, wherein the compound is therapeutic to the female reproductive cells.

64. The method of claim 61, wherein the compound modulates implantation of a fertilized cell into the uterine tissue.

65. The method of claim 61, wherein the compound modulates contraction of the uterus cells.