School of Basic Medical Sciences, State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center and the MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100191, China.
Shanghai Public Health Clinical Center, Shanghai 201508, China.
Science. 2019 Apr 26;364(6438):399-402. doi: 10.1126/science.aau7307.
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.
体外维持终末分化细胞，尤其是肝细胞，一直以来都极具挑战性。在此，我们通过组合五种化学物质（5C）对细胞信号通路进行调节，从而实现了原代人肝细胞（PHH）的长期体外维持。5C 培养的 PHH 表现出类似于新鲜分离细胞的整体基因表达谱和肝细胞特异性功能。此外，这些细胞在 4 周内有效地再现了乙型肝炎病毒（HBV）感染的整个过程，产生了感染性病毒颗粒并形成了 HBV 共价闭合环状 DNA。我们的研究表明，通过化学方法维持 PHH 的功能可支持 HBV 在体外的长期感染，为研究 HBV 细胞生物学和抗病毒药物筛选提供了一个高效的平台。
