Large scale production of retinal pigment epithelial cells
AssigneeHadasit Medical Research Services and Development Ltd.
InventorsBenjamin Eithan Reubinoff, Orna Singer
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Abstract
A method of generating retinal pigment epithelial (RPE) cells is disclosed. The method comprises: (a) culturing human pluripotent stem cells in a human feeder cell-conditioned medium to obtain a cultured population of human pluripotent stem cells; (b) culturing said cultured population of human pluripotent stem cells in a medium comprising a differentiating agent to obtain differentiating cells; and (c) culturing said differentiating cells in a medium comprising one or more members of the TGFβ superfamily.
Claims27
A method of production of retinal pigment epithelial (RPE) cells comprising:
- (a) culturing human pluripotent stem cells in a human feeder cell-conditioned medium in the absence of feeder cells on a non-adherent surface to obtain a cultured population of human pluripotent stem cells, wherein the feeder cell-conditioned medium is isolated from the feeder cells which are used to generate said feeder cell-conditioned medium;
- (b) differentiating said cultured population of human pluripotent stem cells in a medium comprising a differentiating agent to obtain differentiating cells, wherein the differentiating agent is selected from small molecules and proteins;
- and
- (c) further differentiating said differentiating cells in a medium comprising one or more members of the TGFβ superfamily, wherein a yield of between about 50×106 to about 150×106 RPE cells per 175 mL container is capable of being or can be obtained.
A method of increasing the yield of RPE cells comprising:
- (a) culturing human pluripotent stem cells in a human feeder cell-conditioned medium in the absence of feeder cells on a non-adherent surface to obtain a cultured population of human pluripotent stem cells, wherein the feeder cell-conditioned medium is isolated from the feeder cells which are used to generate said feeder cell-conditioned medium;
- (b) differentiating said cultured population of human pluripotent stem cells in a medium comprising a differentiating agent to obtain differentiating cells, wherein the differentiating agent is selected from small molecules and proteins;
- (c) further differentiating said differentiating cells in a medium comprising one or more members of the TGFβ superfamily to obtain RPE cells, wherein a yield of RPE cells obtained is increased by between about 2 to 10 fold compared to a yield of RPE cells that were not cultured in feeder cell-conditioned medium.
A method of treating a retinal disease or disorder comprising:
- (a) culturing human pluripotent stem cells in a human feeder cell-conditioned medium in the absence of feeder cells on a non-adherent surface to obtain a cultured population of human pluripotent stem cells, wherein the feeder cell-conditioned medium is isolated from the feeder cells which are used to generate said feeder cell-conditioned medium;
- (b) differentiating said cultured population of human pluripotent stem cells in a medium comprising a differentiating agent to obtain differentiating cells, wherein the differentiating agent is selected from small molecules and proteins;
- (c) further differentiating said differentiating cells in a medium comprising one or more members of the TGFβ superfamily to obtain RPE cells, wherein a yield of RPE cells obtained is increased by between about 2 to 10 fold compared to a yield of RPE cells that were not cultured in feeder cell-conditioned medium;
- (d) transplanting said RPE cells into the subject's eye, thereby treating the disease or disorder.
Dependencies are inferred from the claim text.