Low oxygen culture conditions for maintaining retinal progenitor cell multipotency
The present invention relates to methods for culturing human retinal progenitor cells under low oxygen conditions to allow the cells to retain the ability to differentiate into photoreceptors following transplantation. The described methods provide cells that can treat a number of ocular diseases, including retinitis pigmentosa and age-related macular degeneration.
1. An isolated substantially homogenous population of multipotent retinal cells produced by a method comprising:
culturing an established cell culture comprising isolated retinal multipotent progenitor cells under low oxygen conditions of from about 1% to about 6% oxygen content of the culture medium and for an effective amount of time while maintaining multipotency of said cells to provide a substantially homogenous population of multipotent retinal progenitor cells, wherein said retinal progenitor cells express activated HIF1 alpha and/or activated HIF2 alpha.
2. The population of claim 1 , wherein the multipotent retinal cells are selected from the group consisting of mouse, rat, simian and human.
3. A composition comprising the cell population of claim 1 and a pharmaceutically acceptable excipient or carrier.
4. The composition of claim 3 , wherein the carrier is a biocompatible matrix.
5. A method for replacing or repairing photoreceptor cells in a patient in need of such treatment comprising administering to said patient an effective amount of the cell population of claim 1 , thereby replacing or repairing photoreceptor cells in said patient.
6. A method for treating or alleviating the symptoms of retinitis pigmentosa in a patient in need of said treatment, comprising administering to said patient an effective amount of the cell population of claim 1 , thereby treating or alleviating the symptoms of retinitis pigmentosa in said patient.
7. A method for treating or alleviating the symptoms of age related macular degeneration in a patient in need of said treatment, comprising administering to said patient an effective amount of the cell population of claim 1 , thereby treating or alleviating the symptoms of age related macular degeneration in said patient.
8. The population of claim 1 , wherein said retinal progenitor cells express both HIF1 alpha and HIF2 alpha.
9. The population of claim 1 , wherein the method further comprises culturing said cells in the presence of at least one exogenous growth factor.
10. The population of claim 1 , wherein the effective amount of time comprises at least 10 passages in the low oxygen conditions.
11. The population of claim 1 , wherein the oxygen content of the culture medium is from about 2% to about 4%.
12. The population of claim 1 , wherein the retinal progenitor cells are obtained from post-natal retinal tissue.
13. The population of claim 1 , wherein the retinal progenitor cells are obtained from the retinal neurosphere.
14. The population of claim 1 , wherein more than about 60% of the cells are of the same or similar phenotype.