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US 10,307,444 B2Granted
Photoreceptors and photoreceptor progenitors produced from pluripotent stem cells
AssigneeAstellas Institute for Regenerative Medicine
InventorsRobert P. Lanza, Shi-Jiang Lu, Wei Wang
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Abstract
Methods are provided for the production of photoreceptor cells and photoreceptor progenitor cells from pluripotent stem cells. Additionally provided are compositions of photoreceptor cells and photoreceptor cells, as well as methods for the therapeutic use thereof. Exemplary methods may produce substantially pure cultures of photoreceptor cells and/or photoreceptor cells.
Claims21
A method of producing photoreceptor progenitor cells, comprising
- (a) culturing pluripotent stem cells in the presence of a BMP signaling inhibitor to produce eye field progenitor cells;
- (b) culturing the eye field progenitor cells under low adherence or non-adherent conditions in a neural differentiation medium in the absence of the BMP signaling inhibitor to form cell spheres;
- (c) culturing the cell spheres under adherent conditions in the neural differentiation medium in the absence of the BMP signaling inhibitor to form attached cells;
- (d) dissociating the attached cells and culturing the cells under low adherence or non-adherent conditions in the neural differentiation medium in the absence of the BMP signaling inhibitor to form further cell spheres;
- (e) culturing the further cell spheres under adherent conditions in the neural differentiation medium in the absence of the BMP signaling inhibitor to form further attached cells;
- (f) repeating steps (d) and (e) at least one additional time;
- and
- thereby obtaining a preparation of photoreceptor progenitor cells, wherein 75%-100% of the photoreceptor progenitor cells in the preparation are PAX6(+) and CHX10(−) at the mRNA level.
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