Novel long-term three-dimensional culture system

• US 20040151729A1
• Filed: 11/04/2003
• Published: 08/05/2004
• Est. Priority Date: 10/28/2002
• Status: Abandoned Application

• Current Assignee: George Michalopoulos, William C. Bowen Jr
• Original Assignee: George Michalopoulos, William C. Bowen Jr
• Inventors: Michalopoulos, George, Bowen, William C. Jr.
• Application Number:
  US10/701,390
• Publication Number:
  US 20040151729A1
• Time in Patent Office: Days
• US Class Current: 424/184.1
• CPC Class Codes: A01N 1/0226 Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
• 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 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/12 Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0671 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

The present invention relates to long-term, three-dimensional cultures of cells in a horizontally-rotating vessel and methods for producing the cultures and using the cultures as explants to treat various conditions.

Claims

1. A method for generation cultures of small tissues comprising incubating cells from said small tissues in a horizontally-rotating vessel, wherein said vessel

(a) contains one or more growth agents that support the growth of said cells and

(b) is coated with extracellular matrix protein that promotes cell adhesion under conditions sufficient to allow for proliferation of said cells while retaining the function of said cells.

2. The method of claim 1 wherein said extracellular matrix protein is type I collagen.

3. The method of claim 1 further comprising culturing said cells in the presence of dexamethasone.

4. The method of claim 1 wherein said small tissue is adrenal tissue.

5. The method of claim 4 wherein said growth agents is transforming growth factor-

6. The method of claim 1 wherein said small tissue is bile duct epithelia.

7. The method of claim 6 wherein said growth agents are hepatocyte growth factor and epidermal growth factor.

8. The method of claim 1 wherein said small tissue is corneal tissue.

9. The method of claim 8 wherein said growth agent is epidermal growth factor.

10. The method of claim 1 wherein said small tissue is pituitary tissue.

11. The method of claim 10 said growth agent is growth hormone.

12. The method of claim 1 wherein said small tissue is thyroid tissue.

13. The method of claim 12 wherein said growth agent is from hypothalamic and pituitary extracts.

14. Small tissue culture derived from a method comprising incubating cells from said small tissues in a horizontally-rotating vessel, wherein said vessel

(a) contains one or more growth agents that support the growth of said cells and

(b) is coated with extracellular matrix protein that promotes cell adhesion and under conditions sufficient to allow for proliferation of said cells while retaining the function of said cells.

15. The small tissue culture of claim 14 wherein said extracellular matrix proteins is type I collagen.

16. The small tissue culture of claim 14 further comprising culturing said cells in the presence of dexamethasone.

17. The small tissue culture of claim 14 wherein said small tissue is adrenal tissue.

18. The small tissue culture of claim 17 wherein said growth agent is transforming growth factor β

.

19. The small tissue culture of claim 14 wherein said small tissue is bile duct epithelia.

20. The small tissue culture of claim 19 wherein said growth agents are hepatocyte growth factor and epidermal growth factor.

21. The small tissue culture of claim 14 wherein said small tissue is corneal epithelia.

22. The small tissue culture of claim 21 wherein said growth agent is epidermal growth factor.

23. The small tissue culture of claim 14 wherein said small tissue is pituitary tissue.

24. The small tissue culture of claim 23 wherein said growth agent is growth hormone.

25. The small tissue culture of claim 14 wherein said small tissue is thyroid tissue.

26. The small tissue culture of claim 25 wherein said growth agents are from hypothalamic and pituitary extracts.

27. A method of providing small tissue function to a subject having a disorder of the small tissue comprising administering to said subject the small tissue culture of claim 14.

28. The method of claim 27 wherein the small tissue is adrenal tissue.

29. The method of claim 27 wherein the small tissue is bile duct epithelia.

30. The method of claim 27 wherein the small tissue is corneal.

31. The method of claim 27 wherein the small tissue is pituitary tissue.

32. The method of claim 27 wherein the small tissue is thyroid tissue.