Long-term functional maintenance of primary human hepatocytes in vitro

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Science  26 Apr 2019:
Vol. 364, Issue 6438, pp. 399-402
DOI: 10.1126/science.aau7307

Keep hepatocytes fresh

Freshly isolated primary human hepatocytes (PHHs) quickly lose their identity and function in vitro, limiting their application in modeling infectious liver diseases and screening drugs. Xiang et al. describe a simple and effective five-chemical culture condition that can maintain the mature function of cultured PHHs for an extended period of time. PHHs cultured in this way can support the entire viral life cycle and recapitulate long-term infection of the hepatitis B virus. This system created a useful drug-screening platform for developing new antiviral strategies.

Science, this issue p. 399


The maintenance of terminally differentiated cells, especially hepatocytes, in vitro has proven challenging. Here we demonstrated the long-term in vitro maintenance of primary human hepatocytes (PHHs) by modulating cell signaling pathways with a combination of five chemicals (5C). 5C-cultured PHHs showed global gene expression profiles and hepatocyte-specific functions resembling those of freshly isolated counterparts. Furthermore, these cells efficiently recapitulated the entire course of hepatitis B virus (HBV) infection over 4 weeks with the production of infectious viral particles and formation of HBV covalently closed circular DNA. Our study demonstrates that, with a chemical approach, functional maintenance of PHHs supports long-term HBV infection in vitro, providing an efficient platform for investigating HBV cell biology and antiviral drug screening.

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