Methods of producing rpe cells and compositions of rpe cells

Abstract

The present invention provides improved methods for producing RPE cells from human embryonic stem cells or from other human pluripotent stem cells. The invention also relates to human retinal pigmented epithelial cells derived from human embryonic stem cells or other human multipotent or pluripotent stem cells. hRPE cells derived from embryonic stem cells am molecularly distinct from adult and fetal-derived RPE cells, and are also distinct from embryonic stem cells. The hRPE cells described hemin are useful for treating retinal degenerative diseases.

1 - 134 . (canceled) 
     
     
         135 . A pharmaceutical preparation comprising
 human RPE cells derived in vitro from human pluripotent stem cells, wherein at least 95% of cells in the preparation are human RPE cells that express three or more of the following markers: RPE-65, Bestrophin, PEDF, CRALBP, Otx2, and Mit-F.   
     
     
         136 . The pharmaceutical preparation of  claim 135 , formulated for administration to a subretinal space of a human eye. 
     
     
         137 . The pharmaceutical preparation of  claim 135 , formulated as a suspension, matrix, or substrate. 
     
     
         138 . The pharmaceutical preparation of  claim 135 , wherein the pharmaceutical preparation comprises at least 10 4  human RPE cells, or at least 10 5  human RPE cells, or at least 10 6  human RPE cells. 
     
     
         139 . The pharmaceutical preparation of  claim 135 , wherein the human RPE cells are derived in vitro from human embryonic stem cells. 
     
     
         140 . The pharmaceutical preparation of  claim 135 , wherein the human RPE cells are derived in vitro from human induced pluripotent stem cells. 
     
     
         141 . The pharmaceutical preparation of  claim 135 , wherein at least 95% of the cells in the preparation are human RPE cells that express 4, 5 or 6 of the markers. 
     
     
         142 . The pharmaceutical preparation of  claim 135 , wherein 99% of the cells in the preparation are human RPE cells that express three or more of the markers. 
     
     
         143 . A method of treating a subject having a condition characterized by retinal degeneration, comprising
 administering to a subject in need thereof an effective amount of a preparation of human RPE cells, wherein the human RPE cells are derived in vitro from human pluripotent stem cells, and wherein at least 95% of the cells in the preparation are human RPE cells that express three or more of the following markers: RPE-65, Bestrophin, PEDF, CRALBP, Otx2, and Mit-F.   
     
     
         144 . The method of  claim 143 , wherein the condition characterized by retinal degeneration is selected from the group consisting of: Stargardt's macular dystrophy, age related macular degeneration, and retinitis pigmentosa, and optionally wherein the preparation is
 (a) formulated as a suspension, matrix, or substrate, and/or   (b) administered by injection into a subretinal space of an eye of the subject.   
     
     
         145 . The method of  claim 143 , wherein the effective amount of the preparation comprises about 10 4  to about 10 6  human RPE cells. 
     
     
         146 . The method of  claim 143 , wherein the human RPE cells are derived in vitro from human embryonic stem cells. 
     
     
         147 . The method of  claim 143 , wherein the human RPE cells are derived in vitro from human induced pluripotent stem cells. 
     
     
         148 . The method of  claim 143 , wherein at least 95% of the cells in the preparation are human RPE cells that express 4, 5 or 6 of the markers. 
     
     
         149 . The method of  claim 143 , wherein 99% of the cells in the preparation are human RPE cells that express three or more of the markers. 
     
     
         150 . The method of  claim 143 , wherein treatment is determined by measuring electroretinogram responses, optomotor acuity threshold, or luminance threshold in the subject. 
     
     
         151 . The method of  claim 143 , wherein the retinal degeneration is associated with photoreceptor loss. 
     
     
         152 . The method of  claim 143 , wherein the human subject maintains photoreceptor thickness following administration of the preparation of human RPE cells. 
     
     
         153 . The method of  claim 152 , wherein the photoreceptor thickness is about 4-5 cells thick. 
     
     
         154 . The method of  claim 143 , wherein the subject has viable endogenous host RPE cells prior to administration of the preparation of human RPE cells.