Back to results
US 2026/0125650 A1Published application
Methods and compositions for generating hepatocytes
AssigneeTrailhead Biosystems Inc.
InventorsMichael A. BUKYS, Xiaoguang FANG, Faruk SENTURK
Life of this patent
Estimating the term…
Abstract
Methods for generating hepatocytes from induced pluripotent stem cells (iPSCs) are provided using chemically-defined culture media in a four-stage culture protocol. Culture media and isolated cell populations are also provided.
Claims14
A method of generating a population of hepatocytes from induced pluripotent stem cells (iPSCs) comprising:
- (a) culturing iPSCs in a culture medium comprising a glycogen synthase kinase 3 beta (GSK3B) inhibitor for 12-120 hours to obtain a first population of endodermal cells;
- (b) culturing the first population of endodermal cells from step (a) in a culture medium lacking a GSK33 inhibitor for 12-120 hours to obtain a second population of endodermal cells;
- (c) culturing the second population of endodermal cells from step (b) in a culture medium comprising a histone deacetylase (HDAC) inhibitor, a retinoic acid (RA) agonist, and a fibroblast growth factor receptor (FGFR;
- e.g., FGFR1) agonist for 24-240 hours to obtain a population of midgut cells;
- (d) culturing the population of midgut cells in a culture medium comprising a glucocorticoid, a cAMP response element-binding protein (CREB) agonist, an antioxidant, a Wnt antagonist, a transforming growth factor beta (TGF-β) antagonist, and a hepatocyte growth factor (HGF) agonist for at least 48 hours to obtain a population of hepatoblasts;
- and
- (e) culturing the population of hepatoblasts in a culture medium comprising an HDAC inhibitor, a glucocorticoid, a CREB agonist, an antioxidant, a Wnt antagonist, a TGF-β antagonist, an HGF agonist, a Notch antagonist, a bone morphogenetic protein (BMP) antagonist, and a hepatocyte function enhancer for at least 48 hours to obtain a population of hepatocytes.
4 more elements
-53. (canceled)
A culture medium for obtaining:
- (a) endodermal cells, wherein the culture medium comprises a GSK3β inhibitor;
- (b) midgut cells, wherein the culture medium comprises an HDAC inhibitor, an RA agonist, and an FGFR (e.g., FGFR1) agonist;
- (c) hepatoblasts, wherein the culture medium comprises a glucocorticoid, a CREB agonist, an antioxidant, a Wnt antagonist, a TGF-β antagonist, and an HGF agonist;
- (d) hepatocytes, wherein the culture medium comprises an HDAC inhibitor, a glucocorticoid, a CREB agonist, an antioxidant, a Wnt antagonist, a TGF-β antagonist, an HGF agonist, a Notch antagonist, a BMP antagonist, and a hepatocyte function enhancer.
1 more element
-57. (canceled)
An isolated cell culture comprising:
- (a) endodermal cells in a culture medium comprising a GSK3β inhibitor;
- (b) midgut cells in a culture medium comprising an HDAC inhibitor, an RA agonist, and an FGFR (e.g., FGFR1) agonist;
- (c) hepatoblasts in a culture medium comprising a glucocorticoid, a CREB agonist, an antioxidant, a Wnt antagonist, a TGF-β antagonist, and an HGF agonist;
- or
- (d) 50 million to 50 billion hepatocytes in a culture medium comprising an HDAC inhibitor, a glucocorticoid, a CREB agonist, an antioxidant, a Wnt antagonist, a TGF-β antagonist, an HGF agonist, a Notch antagonist, a BMP antagonist, and a hepatocyte function enhancer.
2 more elements
-61. (canceled)
Dependencies are inferred from the claim text.