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US 2016/0102289 A1Published application
Generation of keratinocytes from pluripotent stem cells and maintenance of keratinocyte cultures
AssigneeCellular Dynamics International, Inc.
InventorsJunying YU, Ying ZHANG, Lisa HARMS
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Abstract
Provided herein are methods for the generation of functional keratinocyte stem cells that are differentiated directly from human ESCs/iPSCs in a chemically defined serum-free cell culture system, as well as cells derived therefrom and methods of use thereof. Also provided are methods for culturing primary keratinocytes.
Claims45
A method for providing engraftable keratinocyte stem cells by differentiation of pluripotent stem cells comprising:
- (a) forming aggregates of the pluripotent stem cells in a suspension culture in the presence of a defined basal medium;
- (b) culturing the aggregates in a suspension culture in the presence of an initiation culture medium comprising retinoic acid and BMP4 to effect the formation of initiated aggregates;
- (c) culturing the initiated aggregates in a keratinocyte progenitor culture medium comprising cholera toxin and a TGFβR1 kinase inhibitor to effect the formation of a cell population comprising keratinocyte progenitors;
- and
- (d) culturing the keratinocyte progenitors in a keratinocyte stem cell maturation medium to effect the formation of a cell population comprising engraftable keratinocyte stem cells.
2 more elements
A defined keratinocyte culture medium comprising niacinamide and BMP4.
Dependencies are inferred from the claim text.