Compositions and methods for long term culture of hepatocytes
AssigneePeking University
InventorsHongkui Deng, Chengang Xiang, Yuanyuan Du
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Abstract
Provided are compositions for long-term maintenance of functional hepatocytes in culture, a method for improved maintenance of functional hepatocytes in vitro, and functional hepatocytes cultures according to the methods. The culture compositions include at least: one activator of adenylate cyclase, one TGFβinhibitor, one Notch inhibitor, one Wnt inhibitor, and/or one BMP inhibitor. The combinations of compounds are added to any hepatocyte cell culture medium in an effective amount to maintain functional hepatocyte function in vitro, long term. The hepatocytes can be used for in vitro drug research and to model liver disease.
Claims17
A supplemented primary hepatocyte cell culture composition comprising primary hepatocyte cells, and a combination of an activator of adenylate cyclase and a TGFβ inhibitor, in an effective amount to maintain functional hepatocyte function in vitro, for a long term of at least two week, measured as an increase in expression of at least one hepatocyte marker following at least two weeks of in vitro hepatocyte culture in the supplemented cell culture composition, compared to hepatocytes cultured in an unsupplemented culture composition without an activator of adenylate cyclase and a TGFβ inhibitor, wherein the primary hepatocyte cells express the hepatocyte marker at a level that is comparable or more to a level expressed by freshly isolated hepatocytes from the same organism.
Dependencies are inferred from the claim text.