Phenotype profile of human retinal progenitor cells
The present invention relates to substantially homogenous populations of human retinal progenitor cells having the following positive surface markers: SSEA4, CD73, PTK7 and PSA-NCAM. The invention also relates to method for preparing such substantially homogeneous cell populations from human tissue using cell sorting techniques.
1. A composition comprising a pharmaceutically acceptable carrier and a purified population of human retinal progenitor cells having been cultured in from about 10 ng/ml to about 20 ng/ml epidermal growth factor and from about 10 ng/ml to about 20 ng/ml basic fibroblast growth factor in low oxygen conditions consisting of about 3% oxygen for at least 6 hours, said population of human retinal progenitor cells expressing the following positive identifiable markers: SSEA4, CD73, PTK7, PSA-NCAM, CD24, Crx, and Nrl but not expressing CD133, wherein at least 60% of the population of retinal progenitor cells express Ki67 and Cyclin D1, and wherein the pharmaceutically acceptable carrier is suitable for implantation in vivo.
2. The composition of claim 1 wherein the retinal progenitor cells further express one or more of recoverin, Sox2 and Pax6 as a positive identifiable marker.
3. The composition of claim 1 or 2 , wherein the retinal progenitor cells do not express one or more of a negative glial progenitor surface markers A2B5 or CD38.
4. The composition of claim 1 , wherein the population comprises at least about 50% of the retinal progenitor cells express the following positive identifiable markers: SSEA4, Crx, and Nrl.
5. The composition of claim 1 , wherein the population comprises at least about 80% of the retinal progenitor cells express the following positive identifiable markers: SSEA4, and Crx; and wherein at least about 60% of the population of retinal progenitor cells express Nrl.
6. The composition of claim 1 , wherein the pharmaceutically acceptable carrier is a gel, a cell scaffold, or a tube sheet.
7. The composition of claim 1 , wherein at least 80% of the retinal progenitor cells express Cyclin D1 and Ki67.
8. The composition of claim 1 , wherein the purified population of human retinal progenitor cells are isolated by screening for said markers.
9. The composition of claim 8 , wherein the screening comprises flow cytometry.
10. The composition of claim 8 , wherein the screening comprises fluorescence activated cell sorting and antibodies that recognize and bind to the positive cell surface markers.
11. A composition comprising a purified population of human retinal progenitor cells and a fibronectin-coated surface, said human retinal progenitor cells having been cultured in from about 10 ng/ml to about 20 ng/ml epidermal growth factor and from about 10 ng/ml to about 20 ng/ml basic fibroblast growth factor in low oxygen conditions consisting of about 3% oxygen for at least 6 hours, and expressing the following positive identifiable markers: SSEA4, CD73, PTK7, PSA-NCAM, CD24, Crx, and Nrl but not expressing CD133, wherein at least 60% of the population of the retinal progenitor cells express Ki67 and Cyclin D1, and wherein the fibronectin-coated surface is suitable for implantation in vivo.
12. The composition of claim 11 , further comprising an effective amount of stem cell culture media comprising from about 10 ng/ml to about 20 ng/ml epidermal growth factor and from about 10 ng/ml to about 20 ng/ml basic fibroblast growth factor.
13. The composition of claim 11 , wherein at least 80% of the retinal progenitor cells express Cyclin D1 and Ki67.
14. 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 one or more of the population of claim 1 , thereby replacing or repairing photoreceptor cells in said patient.
15. 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 one or more of the population of claim 1 , thereby treating or alleviating the symptoms of retinitis pigmentosa in said patient.
16. A method of 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 one or more of the population of claim 1 , thereby treating or alleviating the symptoms of age related macular degeneration in said patient.