Compositions and methods for long term culture of hepatocytes

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 TGFbeta inhibitor, 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.

Description

technical field [0001] The present invention generally relates to long-term maintenance of hepatocytes and functional maturation of hepatocytes. Background technique [0002] Terminally differentiated cells are functionally stable in vivo, a state that is highly dependent on the precise spatiotemporal regulation of microenvironmental signals. Once isolated, cells undergo microenvironmental changes that lead to functional degradation. Long-term maintenance of terminally differentiated cells in vitro has proven challenging. Once isolated, these cells often lose their specific functions (Haenseler et al., StemCell Reports 8, 1727-1742 (2017); Louch et al., J Mol Cell Cardiol 51, 288-298 (2011); Ootani et al., Nat Med 15, 701-706 (2009 ); Shulman et al., Methods Mol Biol 945, 287-302 (2013). In particular, primary human hepatocytes (PHH) cannot be maintained long-term, limiting their usefulness in modeling hepatotropic infection models such as HBV infection, a global epidemic ...

AI Technical Summary

Problems solved by technology

One of the main reasons for the functional immaturity of hPSC-derived cells is the lack of available culture conditions that allow them to capture the final stages of differentiation

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