Method for reproducible differentiation of clinical-grade retinal pigment epithelium cells
AssigneesThe United States of America, as represented by the Secretary, Department of Health and Human Serv, Cellular Dynamics International, Inc.
InventorsKapil Bharti, Lucas Chase, Feng Xuezhu, Balendu Shekhar Jha
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Abstract
Provided herein are methods of producing an RPE cell population from a starting cell suspension, such as a single cell suspension, of pluripotent stem cells (PSCs). Such a method may comprise culturing the starting single cell suspension of PSCs in differentiation media to produce human RPE cells.
Claims26
A method for producing human retinal pigment epithelial (RPE) cells, comprising:
- a) obtaining a starting population comprising human induced pluripotent stem cells (iPSCs) that are dissociated into essentially single cells;
- b) culturing the iPSCs in a retinal induction medium to initiate differentiation of the cells into retinal lineage cells;
- c) further culturing the retinal lineage cells in a retinal differentiation medium to further differentiate the retinal lineage cells;
- d) culturing the cells in retinal medium to form differentiating RPE cells;
- and
- e) culturing the RPE cells in a RPE maturation medium, thereby producing human RPE cells;
- wherein the method does not comprise the formation of embryoid bodies.
16-18. (canceled)
A method for producing human retinal pigment epithelial (RPE) cells, comprising:
- a) obtaining a starting population comprising human induced pluripotent stem cells (iPSCs) that are dissociated into essentially single cells in a fully defined medium;
- b) culturing the iPSCs on laminin, vitronectin or a combination thereof, in a retinal induction medium comprising LDN193189, CKI-7, and SB431542 to initiate differentiation of the cells into retinal lineage cells;
- c) further culturing the retinal lineage cells in a retinal differentiation medium comprising LDN193189, CKI-7, SB431542, and PD0325901 to further differentiate the retinal lineage cells;
- d) culturing the cells in retinal medium comprising nicotinamide and Activin A to form differentiating RPE cells;
- and
- e) culturing the RPE cells in a RPE maturation medium, thereby producing human RPE cells;
- wherein the method does not comprise the formation of embryoid bodies.
Dependencies are inferred from the claim text.