Modalities for the treatment of degenerative diseases of the retina

1. A method of producing isolated human RPE cells, comprising:

(a) culturing a multilayer culture of human pluripotent cells that express Oct-4, alkaline phosphatase, SSEA-3, SSEA-4, TRA-I-60, and TRA-I-81 under adherent conditions for a sufficient time for the appearance of a pigmented cell population having a cobblestone, polygonal, epithelial-like appearance and comprising cells that express RPE65 and bestrophin; and

(b) isolating the pigmented cell population from the resultant cell culture, thereby obtaining human RPE cells.

2. The method of claim 1 , wherein the multilayer culture of step (a) is produced by allowing pluripotent cells to overgrow and form a thick multilayer of cells.

3. The method of claim 1 , wherein the multilayer culture of step (a) is produced by allowing pluripotent cells to overgrow and form a thick multilayer of cells on a substrate comprising laminin, fibronectin, collagen I, collagen IV, or MATRIGEL (TM) a soluble preparation from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells.

4. A method of producing isolated human RPE cells, comprising:

(a) culturing embryoid bodies comprising human pluripotent cells that express Oct-4, alkaline phosphatase, SSEA-3, SSEA-4, TRA-I-60, and TRA-I-81for a sufficient time for the appearance of a pigmented cell population having a cobblestone polygonal epithelial-like appearance and comprising cells that express RPE65 and bestrophin; and

(b) isolating the pigmented cell population from the resultant cell culture, thereby obtaining human RPE cells.

5. The method of claim 4 , wherein said embryoid bodies are produced by culturing pluripotent cells under low adherent conditions.

6. A method of producing isolated human RPE cells, comprising:

(a) differentiating in vitro human pluripotent cells that express Oct-4, alkaline phosphatase,SSEA-3, SSEA-4, TRA-I-60, and TRA-I-81to obtain a culture of a pigmented cell population having a cobblestone polygonal epithelial-like appearance and comprising cells that express RPE65 and bestrophin; and

(b) isolating the pigmented cell population from the resultant cell culture, thereby obtaining human RPE cells.

7. The method of claim 6 , further comprising cryopreserving said human RPE cells.

8. The method of claim 7 , wherein said cryopreserved human RPE cells are suitable for prevention or treatment of a retinal disease or condition in a human subject in need thereof.

9. The method of claim 6 , wherein said human RPE cells are suitable for prevention or treatment of a retinal disease or condition in a human subject in need thereof.

10. The method of claim 9 , wherein said human RPE cells are disposed on a substrate or matrix.

11. The method of claim 10 , wherein said substrate or matrix comprises at least one of gelatin, fibronectin, laminin, collagen, or extracellular matrix.

12. The method of claim 6 , wherein said human RPE cells are suitable for prevention or treatment of a retinal disease or condition comprising retinitis pigmentosa, RPE detachment, retinal dysplasia, retinal atrophy, retinopathy, macular dystrophy, cone dystrophy, cone-rod dystrophy, Malattia Leventinese, Doyne honeycomb dystrophy, Sorsby's dystrophy, Stargardt disease, pattern/butterfly dystrophies, Best vitelliform dystrophy, North Carolina dystrophy, central areolar choroidal dystrophy, angioid streaks, or a toxic maculopathy in a human subject in need thereof.

13. The method of claim 6 , wherein RPE65 expression is detected by RT-PCR.

14. The method of claim 6 , wherein bestrophin expression is detected by Western blot or immunostaining.

15. The method of claim 6 , wherein said human RPE cells are dispersed in a suspension.

16. The method of claim 6 , wherein said human RPE cells are passaged one or more times.