Adipose derived stromal cells exhibiting characteristics of endothelial cells

• US 20060171932A1
• Filed: 01/27/2006
• Published: 08/03/2006
• Est. Priority Date: 01/31/2005
• Status: Abandoned Application

• Current Assignee: Artecel Inc.
• Original Assignee: Cognate Therapeutics, Inc.
• Inventors: Hendricks, James K., Mitchell, James B. II
• Application Number:
  US11/342,096
• Publication Number:
  US 20060171932A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2506/1384 from adipose-derived stem cells [ADSC], from adipose stromal stem cells
• CPC Class Codes: C12N 5/0667 Adipose-derived stem cells [ADSC]; Adipose stromal stem cells
• CPC Class Codes: C12N 5/069 Vascular Endothelial cells

Abstract

The present invention encompasses an adipose-derived adult stromal (ADAS) cell exhibiting at least one characteristic of a pre-endothelial cell and/or an endothelial cell. The present invention also encompasses compositions and methods for generating an adipose-derived adult stromal to exhibit at least one characteristic of a pre-endothelial cell and/or an endothelial cell. Methods for using the cells in vascular transplantation, tissue engineering, regulation of angiogenesis, vasculogenesis, and the treatment of numerous disorders including heart disease are also included.

Claims

1. An isolated adipose tissue-derived adult stromal (ADAS) cell induced to express at least one characteristic of a pre-endothelial cell.

2. The cell of claim 1, wherein said cell is induced to differentiate in vitro.

3. The cell of claim 1, wherein said cell is induced to differentiate in vivo.

4. The cell of claim 1, wherein exogenous genetic material has been introduced into said cell.

5. The cell of claim 1, wherein said cell is derived from a human.

6. The cell of claim 1, wherein said cell expresses at least one of CD34 and CD31.

7. The cell of claim 1, wherein said cell expresses at least one of CD34 and CD31 at a higher level when compared with the expression level of CD34 and CD31, respectively, from an otherwise identical ADAS cell not induced to express at least one characteristic of a pre-endothelial cell.

8. The cell of claim 1, wherein said cell expresses at least one of CD34, CD31, CD40, CD63, or a combination thereof.

9. The cell of claim 1, wherein said cell expresses at least one of CD34, CD31, CD40, CD63, or a combination thereof at a higher level when compared with the expression level of CD34, CD31, CD40 and CD63, respectively, from an otherwise identical ADAS cell not induced to express at least one characteristic of a pre-endothelial cell.

10. A method of differentiating an isolated adipose tissue-derived adult stromal (ADAS) cell to express at least one characteristic of a pre-endothelial cell, the method comprising incubating said cell in MII medium followed by incubating said cell in MIII medium.

11. The method of claim 10, wherein said cell is derived from a human.

12. The method of claim 10, wherein said MII medium comprises N2 supplement, B27 supplement, glutamine and fibroblast growth factor (FGF).

13. The method of claim 12, wherein the concentration of said glutamine is about 2.3 mM.

14. The method of claim 12, wherein the concentration of said FGF is about 10 ng/mL.

15. The method of claim 10, wherein said MIII medium comprises N2 supplement, B27 supplement, glutamine, nicotinamide and fetal bovine serum (FBS).

16. The method of claim 15, wherein the concentration of said glutamine is about 2.3 mM.

17. The method of claim 15, wherein the concentration of said nicotinamide is about 10 mM.

18. The method of claim 15, wherein the concentration of said FBS is about 2%.

19. A differentiation medium for differentiating an isolated adipose tissue-derived adult stromal (ADAS) cell into a cell exhibiting at least one characteristic of a pre-endothelial cell, wherein said medium is supplemented with N2 supplement, B27 supplement, glutamine and fibroblast growth factor (FGF), further wherein said medium is designated as MII medium.

20. The medium of claim 19, wherein the concentration of said glutamine is about 2.3 mM.

21. The medium of claim 19, wherein the concentration of said FGF is about 10 ng/mL.

22. A differentiation medium for differentiating an isolated adipose tissue-derived adult stromal (ADAS) cell into a cell exhibiting at least one characteristic of a pre-endothelial cell, wherein said medium is supplemented with N2 supplement, B27 supplement, glutamine, nicotinamide and fetal bovine serum (FBS), further wherein said medium is designated as MIII medium.

23. The medium of claim 22, wherein the concentration of said glutamine is about 2.3 mM.

24. The medium of claim 22, wherein the concentration of said nicotinamide is about 10 mM.

25. The medium of claim 22, wherein the concentration of said FBS is about 2%.

26. A method of inducing vasculogenesis in an animal comprising:

a) inducing an isolated adipose tissue-derived adult stromal (ADAS) cell to express at least one characteristic of a pre-endothelial cell; and

b) administering said cell so induced into said animal.

27. The method of claim 26, wherein said ADAS cell is isolated from said animal.

28. The method of claim 26, wherein said ADAS cell is isolated from an allogeneic donor.

29. The method of claim 26, wherein said ADAS cell is isolated from a xenogeneic donor.

30. The method of claim 26, wherein said ADAS cell is derived from a human.

31. A method of determining the ability of a compound to affect the differentiation of an isolated adipose tissue-derived adult stromal (ADAS) cell into a pre-endothelial cell and/or endothelial cell, the method comprising:

a) culturing said ADAS cell in a stromal cell medium for a period of time;

b) replacing said stromal cell medium with a differentiation medium comprising a compound or a control vehicle;

c) incubating said ADAS cell in said differentiation medium comprising said compound or said control vehicle for a period of time;

d) determining the number or percentage of differentiated cells using said differentiation medium comprising said compound from step (c);

e) determining the number of percentage of differentiated cells in the cells using said differentiation medium containing said vehicle alone from step (c);

f) comparing the number or percentage of differentiated cells from steps (d) and (e);

g) a greater number of percentage of differentiated cells from step (d) compared to the number of percentage of differentiated cells from step (e) indicates that said compound is capable of inducing differentiation of said ADAS cell into a pre-endothelial cell and/or endothelial cell.

32. The method of claim 31, wherein said cell is derived from a human.