Vascularization enhanced graft constructs

• US 20030216811A1
• Filed: 05/02/2003
• Published: 11/20/2003
• Est. Priority Date: 05/02/2002
• Status: Active Grant

• Current Assignee: Clarian Health Partners, Inc.
• Original Assignee: Clarian Health Partners, Inc.
• Inventors: Badylak, Stephen F.
• Granted Patent: US 7,815,686 B2
• Time in Patent Office: Days
• US Class Current: 623/17.16
• 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: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2502/1347 Smooth muscle cells
• CPC Class Codes: C12N 2502/23 Gastro-intestinal tract cells
• CPC Class Codes: C12N 2502/28 Vascular endothelial cells
• CPC Class Codes: C12N 2509/00 Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 5/069 Vascular Endothelial cells
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691

Abstract

An intestinal submucosa tissue graft construct for use in repairing diseased or damaged tissues is provided. The graft construct comprises vertebrate intestinal submucosa tissue, added endothelial cells, and at least one additional preselected, exogenous population of cells which enhances initiation of the formation of vessel-like structures in the graft. The preselected population of cells can be a population of non-keratinized or keratinized epithelial cells or a population of mesodermally derived cells selected from the group consisting of fibroblasts, smooth muscle cells, skeletal muscle cells, cardiac muscle cells, multi-potential progenitor cells, pericytes, osteogenic cells, and any other suitable cell type, preferably selected based on the tissue to be repaired. Methods for enhancing the vascularization in vivo of these intestinal submucosa tissue graft constructs and for preparing these graft constructs are also provided.

Claims

1. An intestinal submucosa tissue graft construct for use in repairing diseased or damaged tissues, said tissue graft construct comprising vertebrate intestinal submucosa tissue, added endothelial cells, and at least one additional preselected, exogenous population of cells.

2. The graft construct of claim 1 wherein the at least one additional cell population comprises a population cells selected from the group consisting of keratinized epithelial cells, non-keratinized epithelial cells, and mesodermally derived cells.

3. The graft construct of claim 1 wherein the at least one additional cell population comprises a population of smooth muscle cells.

4. The graft construct of claim 1 wherein the at least one additional cell population comprises a population of smooth muscle cell progenitor cells.

5. The graft construct of claim 1 wherein the intestinal submucosa tissue is delaminated from the tunica muscularis layer and at least the luminal portion of the tunica mucosa layer of said intestinal tissue.

6. The graft construct of claim 1 further comprising a heparinase.

7. The graft construct of claim 1 further comprising a growth factor selected from the group consisting of vascular endothelial cell-derived growth factor, platelet-derived growth factor, a platelet-derived growth factor-like molecule, transforming growth factor β

, and a serum growth factor.

8. The graft construct of claim 1 wherein the intestinal submucosa tissue is seeded with the additional preselected population of cells after the intestinal submucosa tissue is seeded with the endothelial cells.

9. The graft construct of claim 1 wherein the intestinal submucosa tissue is seeded with the endothelial cells after the intestinal submucosa tissue is seeded with the additional preselected population of cells.

10. The graft construct of claim 1 wherein the intestinal submucosa tissue is seeded with the endothelial cells and the additional preselected population of cells simultaneously or nearly simultaneously.

49. A method for treating diseased or damaged tissue in a patient, comprising repairing the diseased or damaged tissue with a tissue graft construct according to any of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 42, 43, 45, or 47.

11. A method for enhancing the vascularization in vivo of an intestinal submucosa tissue graft construct, said method comprising the steps of:

seeding said intestinal submucosa tissue in vitro with a population of endothelial cells and at least one additional preselected, exogenous population of cells; and

implanting said graft construct into a vertebrate at a site in need of repair.

12. The method of claim 11 wherein the at least one additional cell population comprises a population of cells selected from the group consisting of keratinized epithelial cells, non-keratinized epithelial cells, and mesodermally derived cells.

13. The method of claim 11 wherein the at least one additional cell population comprises a population of smooth muscle cells.

14. The method of claim 11 wherein the at least one additional cell population comprises a population of smooth muscle cell progenitor cells.

15. The method of claim 11 wherein the intestinal submucosa tissue is delaminated from the tunica muscularis layer and at least the luminal portion of the tunica mucosa layer of said intestinal tissue.

16. The method of claim 11 wherein the graft construct further comprises a heparinase.

17. The method of claim 11 wherein the graft construct further comprises a growth factor selected from the group consisting of vascular endothelial cell-derived growth factor, platelet-derived growth factor, a platelet-derived growth factor-like molecule, transforming growth factor β

and a serum growth factor.

18. The method of claim 11 further comprising the step of culturing said endothelial cells on said intestinal submucosa tissue for a time sufficient to form vessels or vessel-like structures in vitro before implanting said graft construct into the vertebrate.

19. The method of claim 11 wherein the intestinal submucosa tissue is seeded with the additional preselected population of cells after the intestinal submucosa tissue is seeded with the endothelial cells.

20. The method of claim 11 wherein the intestinal submucosa tissue is seeded with the endothelial cells after the intestinal submucosa tissue is seeded with the additional preselected population of cells.

21. The method of claim 11 wherein the intestinal submucosa tissue is seeded with the endothelial cells and the additional preselected population of cells simultaneously or nearly simultaneously.

22. The method of claim 18 wherein the intestinal submucosa tissue is seeded with the additional preselected population of cells after the intestinal submucosa tissue is seeded with the endothelial cells.

23. The method of claim 18 wherein the intestinal submucosa tissue is seeded with the endothelial cells after the intestinal submucosa tissue is seeded with the additional preselected population of cells.

24. The method of claim 18 wherein the intestinal submucosa tissue is seeded simultaneously with the endothelial cells and the additional preselected population of cells.

25. The method of any of claims 19, 20, or 21 wherein neither the endothelial cells nor the additional preselected population of cells is expanded prior to implanting the graft construct.

26. The method of any of claims 19, 20, or 21 further comprising the steps of culturing the endothelial cells on the intestinal submucosa tissue for a time sufficient to expand the endothelial cells without forming vessels or vessel-like structures in vitro and culturing the additional preselected population of cells on the intestinal submucosa tissue for a time sufficient to expand the additional preselected population of cells before implanting the graft construct into the vertebrate.

27. The method of any of claims 19, 20, or 21 further comprising the step of culturing the endothelial cells on the intestinal submucosa tissue for a time sufficient to expand the endothelial cells without forming vessels or vessel-like structures in vitro.

28. The method of claim 27 wherein the additional preselected population of cells is not expanded prior to implanting the graft construct.

29. The method of any of claims 19, 20, or 21 further comprising the step of culturing the additional preselected population of cells on the intestinal submucosa tissue for a time sufficient to expand the preselected population of cells prior to implanting the graft construct.

30. The method of claim 29 wherein the endothelial cells are not expanded prior to implanting the graft construct.

31. The method of any of claims 22, 23, or 24 wherein the additional preselected population of cells is expanded prior to implanting the graft construct.

32. The method of any of claims 22, 23, or 24 wherein the additional preselected population of cells is not expanded prior to implanting the graft construct.

33. A method of preparing an intestinal submucosa tissue graft construct for use in repairing diseased or damaged tissues, said method comprising the step of seeding said intestinal submucosa tissue in vitro with a population of endothelial cells, and at least one additional preselected, exogenous population of cells.

34. The method of claim 33 further comprising the step of culturing said endothelial cells on said intestinal submucosa tissue for a time sufficient to form vessels or vessel-like structures in vitro before implanting said graft construct into the vertebrate.

35. A method for enhancing the vascularization in vivo of an intestinal submucosa tissue graft construct said method comprising the steps of:

seeding said intestinal submucosa tissue in vitro with a population of endothelial cells and at least one additional preselected, exogenous population of cells;

culturing in vitro said endothelial cells on said intestinal submucosa tissue for a time sufficient to induce the formation of vessels or vessel-like structures; and

implanting said graft construct into a vertebrate in a site in need of repair.

36. The method of claim 35 wherein the at least one additional cell population comprises a population of cells selected from the group consisting of keratinized epithelial cells, non-keratinized epithelial cells, and mesodermally derived cells.

37. The method of claim 35 wherein the at least one additional cell population comprises a population of smooth muscle cells.

38. The method of claim 35 wherein the at least one additional cell population comprises a population of smooth muscle cell progenitor cells.

39. The method of claim 35 wherein the intestinal submucosa tissue is seeded with the additional preselected population of cells after the intestinal submucosa tissue is seeded with the endothelial cells.

40. The method of claim 35 wherein the intestinal submucosa tissue is seeded with the endothelial cells after the intestinal submucosa tissue is seeded with the additional preselected population of cells.

41. The method of claim 35 wherein the intestinal submucosa tissue is seeded simultaneously with the endothelial cells and the additional preselected population of cells.

42. A vascularized intestinal submucosa tissue graft construct for use in repairing diseased or damaged tissues, said tissue graft construct comprising vertebrate intestinal submucosa tissue, added endothelial cells, and at least one additional preselected, exogenous population of cells wherein said endothelial cells have been cultured on said intestinal submucosa tissue for a time sufficient to form vessels or vessel-like structures in vitro.

43. An intestinal submucosa tissue graft construct having vessel-like structures for use in repairing diseased or damaged tissues, said tissue graft construct comprising vertebrate intestinal submucosa tissue, added endothelial cells, and at least one additional preselected, exogenous population of cells wherein said additional population of cells enhances the initiation of formation of said vessel-like structures in said graft construct.

44. A method of preparing an intestinal submucosa tissue graft construct having vessel-like structures for use in repairing diseased or damaged tissues, said method comprising the step of seeding said intestinal submucosa tissue in vitro with a population of endothelial cells, and at least one additional preselected, exogenous population of cells wherein said additional population of cells enhances the initiation of formation of said vessel-like structures in said graft construct.

45. A tissue graft construct for use in repairing diseased or damaged tissues, said tissue graft construct comprising submucosa tissue, added endothelial cells, and at least one additional preselected, exogenous population of cells wherein the submucosa tissue is delaminated from the tunica muscularis layer and at least the luminal portion of the tunica mucosa layer of said tissue.

46. A method of preparing a tissue graft construct for use in repairing diseased or damaged tissues, said method comprising the step of seeding in vitro submucosa tissue with a population of endothelial cells, and at least one additional preselected, exogenous population of cells wherein the submucosa tissue is delaminated from the tunica muscularis layer and at least the luminal portion of the tunica mucosa layer of said tissue.

47. A tissue graft construct having vessel-like structures for use in repairing diseased or damaged tissues, said tissue graft construct comprising submucosa tissue, added endothelial cells, and at least one additional preselected, exogenous population of cells wherein said additional population of cells enhances the initiation of formation of said vessel-like structures in said graft construct.

48. A method of preparing a tissue graft construct having vessel-like structures for use in repairing diseased or damaged tissues, said method comprising the step of seeding in vitro submucosa tissue with a population of endothelial cells, and at least one additional preselected, exogenous population of cells wherein said additional population of cells enhances the initiation of formation of said vessel-like structures in said graft construct.