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. A method of expanding multipotent human retinal progenitor cells in vitro and maintaining the multipotency of said cells in vivo, said method comprises the steps of
establishing a cell culture comprising isolated retinal multipotent progenitor cells, and
culturing said 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 an expanded population of multipotent retinal progenitor cells,
wherein said retinal progenitor cells express both express both HIF1 alpha and HIF2 alpha.
2. The method of claim 1 further comprising isolating the expanded population of cells from the culture media.
3. The method of claim 2 further comprising transplanting said cells into a host.
4. The method of claim 3 further comprising measuring said cells cultured under low oxygen conditions for multipotency.
5. The method of claim 1 further comprising culturing said cells in the presence of at least one exogenous growth factor.
6. The method of claim 1 wherein the effective amount of time comprises at least 10 passages in the low oxygen conditions.
7. The method of claim 1 wherein the oxygen content of the culture medium is from about 2% to about 4%.
8. The method of claim 1 where the retinal progenitor cells are obtained from post-natal retinal tissue.
9. The method of claim 1 wherein the retinal progenitor cells are obtained from the retinal neurosphere.