Claims
- 1. A method of stimulating proliferation of adult human corneal endothelial cells, comprising:exposing adult human corneal endothelial cells to an effective amount of at least one growth factor that promotes proliferation of corneal endothelial cells; and subsequently exposing the adult human corneal endothelial cells to an effective amount of at least one agent that promotes interruption of cell-cell contacts between adjacent corneal endothelial cells.
- 2. The method of claim 1, further comprising, subsequent to the cell-cell contact interrupting step, exposing the adult human corneal endothelial cells again to the at least one growth factor.
- 3. The method of claim 1, wherein the adult human corneal endothelial cells are in a form selected from the group consisting of a corneal cell culture, a corneal tissue culture, a corneal organ culture, an intact cornea, and an intact eye.
- 4. The method of claim 1, wherein the at least one agent comprises a calcium chelator.
- 5. The method of claim 4, wherein the calcium chelator is ethylenediaminetetraacetic acid (EDTA) or ethylene glycol-bis[β-aminoethylether]-N,N,N′,N′-tetraacetic acid (EGTA).
- 6. The method of claim 4 or claim 5, wherein the agent is administered to the corneal endothelial cells in a concentration within a range of about 0.02-3.0 mg/ml.
- 7. The method of claim 4 or 5, wherein the agent is administered to the adult human corneal endothelial cells in a concentration within a range of about 0.2-2.0 mg/ml.
- 8. The method of claim 1, wherein the agent is an antibody that specifically binds to a cell surface protein on the corneal adult human corneal endothelial cell that is involved in cell-cell adhesion.
- 9. The method of claim 8, wherein the agent is an antibody that specifically binds to a protein selected from the group consisting of a cadherin, ZO-1 protein, and connexin-43.
- 10. The method of claim 1, wherein the interruption step results in an interruption in cell-cell contacts in at least 15% of the adult human corneal endothelial cells.
- 11. The method of claim 1, wherein the interruption step results in an interruption in cell-cell contacts in at least 50% of the adult human corneal endothelial cells.
- 12. The method of claim 1, wherein the interruption step results in an interruption in cell-cell contacts in at least 80% of the adult human corneal endothelial cells.
CROSS REFERENCE TO RELATED APPLICATIONS
This application claims priority from U.S. Provisional Application No. 60/145,171, filed on Jul. 22, 1999, and International Application No. PCT/US00/03531, entitled GROWTH MEDIUM FOR HUMAN CORNEAL ENDOTHELIAL CELLS, filed on Feb. 11, 2000, which are fully incorporated herein by reference.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
The work leading to the present invention was supported in part by federal funding under grant number NEI RO1 EY05767. Therefore, the U.S. government may have certain rights in this invention.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/US00/40471 |
|
WO |
00 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO01/06843 |
2/1/2001 |
WO |
A |
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
5104787 |
Lindstrom et al. |
Apr 1992 |
A |
Non-Patent Literature Citations (4)
Entry |
Tadashi Senoo, “Stimulation of Corneal Endothelial Cell Proliferation by Interleukins and Complete Mitogens”, Dokkyo Journal of Medical Sciences, vol. 22 pp. 159-170 (1995).* |
Ko-Hua Chen et al., “TGF-β2 in Aqueous Humor Suppresses S-Phase Entry in Cultured Corneal Endothelial Cells”, Investigative Ophthatlmology & Visual Science, vol. 40, No. 11, pp. 2513-2519 (1999). |
Tadashi Senoo, “Stimulation of Corneal Endothelial Cell Proliferation By Interleukins and Complete Mitogens”, Dokkyo Journal of Medical Sciences, vol. 22, pp. 159-170 (1995). |
Nancy C. Joyce et al., “Mitotic Inhibition of Corneal Endothelium in Neonatal Rats”, Investigative Ophthalmology & Visual Science, vol. 39, No. 13, pp. 2572-2583 (1998). |
Provisional Applications (1)
|
Number |
Date |
Country |
|
60/145171 |
Jul 1999 |
US |