Vascularization enhanced graft constructs

• US 8,084,048 B2
• Filed: 09/10/2009
• Issued: 12/27/2011
• Est. Priority Date: 05/02/2002
• Status: Active Grant

• Current Assignee: Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Original Assignee: Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Inventors: Badylak, Stephen F.
• Primary Examiner(s): DAVIS, RUTH A
• Application Number:
  US12/557,176
• Publication Number:
  US 20090324681A1
• Time in Patent Office: 838 Days
• Field of Search: 424/400, 424/422, 424/572, 435/325, 435/371
• US Class Current: 424/400
• CPC Class Codes: A61K 35/34 Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes vascular smooth muscle A61K35/44
• 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 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3808 Endothelial cells
• 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 27/383 Nerve cells, e.g. dendritic cells, Schwann cells
• CPC Class Codes: A61L 27/3886 comprising two or more cell types

Abstract

A tissue graft construct for use in repairing diseased or damaged tissues is provided. The tissue graft construct comprises a matrix composition selected from the group consisting of liver basement membrane and extracts and hydrolysates thereof, and processed collagen from vertebrate non-submucosal sources, added endothelial cells, and at least one additional preselected, exogenous population of cells which enhance the initiation of vessel-like structures in the grant. 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 tissue graft constructs and for preparing these graft constructs are also provided.

Claims

1. A method for enhancing the vascularization in vivo of a tissue graft construct, said method comprising the steps of:

seeding in vitro a matrix composition selected from the group consisting of liver basement membrane and extracts and hydrolysates thereof, and processed collagen from vertebrate non-submucosal sources, with a population of endothelial cells and at least one additional preselected, exogenous population of cells to form the graft construct wherein said additional population of cells enhances the initiation of vascularization of said graft construct; and

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

2. The method of claim 1 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.

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

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

5. The method of claim 1 wherein the at least one additional cell population comprises fibroblasts.

6. The method of claim 1 wherein the graft construct further comprises a heparinase.

7. The method of claim 1 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.

8. The method of claim 1 further comprising the step of culturing said endothelial cells on said matrix composition for a time sufficient to form vessels or vessel-like structures in vitro before implanting said graft construct into the vertebrate.

9. The method of claim 8 wherein the matrix composition is seeded with the additional preselected population of cells after the matrix composition is seeded with the endothelial cells.

10. The method of claim 8 wherein the matrix composition is seeded with the endothelial cells after the matrix composition is seeded with the additional preselected population of cells.

11. The method of claim 8 wherein the matrix composition is seeded simultaneously with the endothelial cells and the additional preselected population of cells.

12. The method of claim 11 wherein the additional preselected population of cells is expanded prior to implanting the graft construct.

13. The method of claim 11 wherein the additional preselected population of cells is not expanded prior to implanting the graft construct.

14. The method of claim 1 wherein the matrix composition is seeded with the additional preselected population of cells after the matrix composition is seeded with the endothelial cells.

15. The method of claim 14 wherein neither the endothelial cells nor the additional preselected population of cells is expanded prior to implanting the graft construct.

16. The method of claim 14 further comprising the steps of culturing the endothelial cells on the matrix composition 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 matrix composition for a time sufficient to expand the additional preselected population of cells before implanting the graft construct into the vertebrate.

17. The method of claim 14 further comprising the step of culturing the endothelial cells on the matrix composition for a time sufficient to expand the endothelial cells without forming vessels or vessel-like structures in vitro.

18. The method of claim 17 wherein the additional preselected population of cells is not expanded prior to implanting the graft construct.

19. The method of claim 14 further comprising the step of culturing the additional preselected population of cells on the matrix composition for a time sufficient to expand the preselected population of cells prior to implanting the graft construct.

20. The method of claim 19 wherein the endothelial cells are not expanded prior to implanting the graft construct.

21. The method of claim 1 wherein the matrix composition is seeded with the endothelial cells after the matrix composition is seeded with the additional preselected population of cells.

22. The method of claim 1 wherein the matrix composition is seeded with the endothelial cells and the additional preselected population of cells simultaneously or nearly simultaneously.