Methods for the production of hepatic stellate cells from human pluripotent stem cells
AssigneeFUJIFILM Holdings America Corporation
InventorsYusuke MORI, Tadanori YAMADA, Ryota HABA, Hayato KOBAYASHI, Setsu ENDOH, Kristin WILLIS, Igor GUREVICH
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Abstract
Provided herein are methods for producing quiescent hepatic stellate cells (HSCs) from pluripotent stem cells and compositions thereof. In particular aspects, the HSCs are produced without a cell sorting or enrichment step and under feeder-free and serum-free conditions.
Claims49
An in vitro method for producing quiescent hepatic stellate cells (HSCs) comprising:
- (a) culturing pluripotent stem cells (PSCs) in a first media;
- (b) further culturing the PSCs in a second media comprising a bone morphogenic protein (BMP) agonist and a glycogen synthase kinase 3 (GKS3) inhibitor, thereby producing mesodermal cells;
- (c) culturing the mesodermal cells in the presence of a survival agent, thereby producing aggregates;
- and
- (d) differentiating the aggregates in a third media comprising a transforming growth factor beta (TFGB) inhibitor and a BMP inhibitor, thereby producing a population of quiescent HSCs.
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A method of producing a cell population comprising at least 20% LRAT-protein expressing cells, wherein the cell population has not undergone a cell sorting step.
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Dependencies are inferred from the claim text.