Chemical differentiation of pluripotent stem cells into retinal epithelial cells

Abstract

The present invention is based in part on a chemically defined method of generating retinal epithelial cells and retinal pigmented epithelial cells from human pluripotent stem cells (hpSCs). The present invention also provides methods and kits for treating degenerative eye disorders.

What is claimed is: 
     
         1 . A method of treating a subject suffering from a degenerative eye disorder comprising administering retinal pigmented epithelial cells to a subject in need thereof. 
     
     
         2 . The method of  claim 1 , wherein the retinal pigmented epithelial cells are produced by a method comprising:
 a) culturing hpSCs on a feeder layer;   b) transferring the hpSCs to feeder layer free culture conditions;   c) culturing the hpSCs with at least one retinal cell inducing agent to produce retinal epithelial progenitor cells; and   d) culturing the retinal epithelial progenitor cells with a GSK-3 inhibitor,   thereby producing retinal pigmented epithelial cells.   
     
     
         3 . The method of  claim 1 , wherein the hpSCs are selected from the group consisting of: embryonic stem cells, parthenogenetic stem cells, induced pluripotent stem cells and nuclear transfer derived stem cells. 
     
     
         4 . The method of  claim 1 , wherein the feeder layer comprises mitomycin-C inactivated mouse embryonic fibroblast cells. 
     
     
         5 . The method of clam  1 , wherein the feeder layer free culture conditions comprise culturing the hpSCs on plates coated with a xeno-free substrate. 
     
     
         6 . The method of  claim 2 , wherein the at least one retinal cell inducing agent is selected from the group consisting of a TGFβR-1 inhibitor, a Wnt inhibitor, a BMP inhibitor and a ROCK inhibitor, or any combination thereof. 
     
     
         7 . The method of  claim 1 , wherein the at least one retinal cell inducing agent comprises SB431542, IWR-1, DMH-1 or Y-27632, or any combination thereof, and the GSK-3 inhibitor is Chir90221. 
     
     
         8 . The method of  claim 1 , wherein the degenerative eye disorder is macular degeneration.