Conversion of Fibroblasts to Hepatocytes In Vitro

  • Pengyu HuangEmail author
  • Lulu Sun
  • Ludi Zhang
  • Lijian Hui
Part of the Methods in Molecular Biology book series (MIMB, volume 1905)


Primary hepatocytes are widely used in regenerative medicine, drug metabolism analysis, and in vitro drug screens. To overcome the shortage of liver donors, several strategies, such as differentiation of pluripotent stem cells and transdifferentiation from somatic cells, were developed to generate hepatocytes from alternative sources. Here, we describe in detail lenti-virus-based procedure for direct conversion of human fibroblasts to hepatocytes (hiHep cells) in vitro. A detailed protocol for preparation of human fibroblasts from scar tissues is also provided. Based on this protocol, FOXA3, HNF1A, and HNF4A are introduced into SV40-large-T-antigen-expressing human scar fibroblasts by lenti-virus. It usually takes about 5–7 days to get epithelial hiHep colonies. SV40-large-T-antigen-expressing hiHep (hiHepLT) cells are proliferative and can be expanded to a large number for potential uses.

Key words

Hepatocytes HiHep cells Transdifferentiation Reprogramming 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Pengyu Huang
    • 1
    Email author
  • Lulu Sun
    • 2
  • Ludi Zhang
    • 2
  • Lijian Hui
    • 1
    • 2
  1. 1.School of Life Science and TechnologyShanghai Tech UniversityShanghaiChina
  2. 2.State Key Laboratory of Cell BiologyCAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of SciencesShanghaiChina

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