Back to results
US 10,047,341 B2Granted
Generation of keratinocytes from pluripotent stem cells and maintenance of keratinocyte cultures
AssigneeFUJIFILM Cellular Dynamics, Inc.
InventorsJunying Yu, Ying Zhang, Lisa Harms
Life of this patent
Estimating the term…
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.
Claims31
A method for providing engraftable human keratinocyte stem cells by differentiation of human 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 bone morphogenetic protein 4 (BMP4) to effect the formation of initiated aggregates;
- (c) culturing the initiated aggregates in a keratinocyte progenitor culture medium comprising cholera toxin and a transforming growth factor beta receptor 1 (TGFBR1) 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 comprising cholera toxin and vascular endothelial growth factor (VEGF) to effect the formation of a cell population comprising engraftable keratinocyte stem cells,
- wherein the keratinocyte stem cells express cytokeratin 15 and CD49f.
3 more elements
Dependencies are inferred from the claim text.