Hepatocyte Expansion Methods
AssigneeKoninklijke Nederlandse Akademie van Wetenschappen
InventorsJohannes Carolus Clevers, Huili Hu
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Abstract
This invention relates to culture methods and media for in vitro expansion of hepatocytes, particularly primary hepatocytes; hepatocyte cultures and organoids obtainable and obtained by said methods; and uses of said hepatocyte cultures and organoids.
Claims21
-56. (canceled)
A composition comprising:
- a. a hepatocyte organoid, wherein the hepatocyte organoid expresses albumin and does not express KRT7 and KRT19;
- and
- b. a hepatocyte culture medium, wherein the hepatocyte culture medium comprises:
- i. 50 to 500 ng/ml of a Fibroblast Growth Factor (FGF) selected from FGF7, FGF10, and a combination thereof;
- ii. a Wnt agonist comprising an R-spondin selected from R-spondin 1, R-spondin 2, R-spondin 3, and R-spondin 4;
- and at least one glycogen synthase kinase 3 (GSK3) inhibitor;
- iii. 5 to 100 ng/ml of epidermal growth factor (EGF);
- iv. 5 to 100 ng/ml of hepatocyte growth factor (HGF);
- and
- v. a transforming growth factor beta (TGF-β) inhibitor, comprising an inhibitor of activin receptor-like kinase ALK4, ALK5 and/or ALK7 signalling pathway.
An in vitro composition consisting essentially of a hepatocyte organoid, wherein the hepatocyte organoid expresses albumin and does not express KRT7 and KRT19.
An in vitro composition comprising a hepatocyte organoid, wherein the hepatocyte organoid expresses albumin and does not express KRT7 and KRT19.
Dependencies are inferred from the claim text.