Susceptibility of chimeric mice with livers repopulated by serially subcultured human hepatocytes to hepatitis B virus

Authors

  • Rie Utoh,

    1. Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization (CLUSTER), Hiroshima Prefectural Institute of Industrial Science and Technology, Hiroshima, Japan
    Current affiliation:
    1. Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan
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  • Chise Tateno,

    1. Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization (CLUSTER), Hiroshima Prefectural Institute of Industrial Science and Technology, Hiroshima, Japan
    2. Hiroshima University Liver Project Research Center, Hiroshima, Japan
    Current affiliation:
    1. PhoenixBio Co., Ltd., Hiroshima, Japan
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  • Chihiro Yamasaki,

    1. Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization (CLUSTER), Hiroshima Prefectural Institute of Industrial Science and Technology, Hiroshima, Japan
    Current affiliation:
    1. PhoenixBio Co., Ltd., Hiroshima, Japan
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  • Nobuhiko Hiraga,

    1. Division of Frontier Medical Science, Department of Medicine and Molecular Science, Programs for Biomedical Research, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan
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  • Miho Kataoka,

    1. Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization (CLUSTER), Hiroshima Prefectural Institute of Industrial Science and Technology, Hiroshima, Japan
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  • Takashi Shimada,

    1. PhoenixBio Co., Ltd., Hiroshima, Japan
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  • Kazuaki Chayama,

    1. Hiroshima University Liver Project Research Center, Hiroshima, Japan
    2. Division of Frontier Medical Science, Department of Medicine and Molecular Science, Programs for Biomedical Research, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan
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  • Katsutoshi Yoshizato

    Corresponding author
    1. Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization (CLUSTER), Hiroshima Prefectural Institute of Industrial Science and Technology, Hiroshima, Japan
    2. Hiroshima University Liver Project Research Center, Hiroshima, Japan
    3. Developmental Biology Laboratory and Hiroshima University 21ststCentury COE Program for Advanced Radiation Casualty Medicine, Department of Biological Science, Graduate School of Science, Hiroshima University, Hiroshima, Japan
    Current affiliation:
    1. PhoenixBio Co., Ltd., Hiroshima, Japan
    • PhoenixBio Co., Ltd., 3-4-1 Kagamiyama, Higashihiroshima, Hiroshima 739-0046, Japan
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    • fax: (81)-82-431-0017


  • Potential conflict of interest: Nothing to report.

Abstract

We previously identified a small population of replicative hepatocytes in long-term cultures of human adult parenchymal hepatocytes (PHs) at a frequency of 0.01%-0.09%. These hepatocytes were able to grow continuously through serial subcultures as colony-forming parenchymal hepatocytes (CFPHs). In the present study, we generated gene expression profiles for cultured CFPHs and found that they expressed cytokeratin 19, CD90 (Thy-1), and CD44, but not mature hepatocyte markers such as tryptophan-2,3-dioxygenase (TO) and glucose-6-phosphatase (G6P), confirming that these cells are hepatic progenitor-like cells. The cultured CFPHs were resistant to infection with human hepatitis B virus (HBV). To examine the growth and differentiation capacity of the cells in vivo, serially subcultured CFPHs were transplanted into the progeny of a cross between albumin promoter/enhancer-driven urokinase plasminogen activator-transgenic mice and severe combined immunodeficient (SCID) mice. The cells were engrafted into the liver and were able to grow for at least 10 weeks, ultimately reaching a maximum occupancy rate of 27%. The CFPHs in the host liver expressed differentiation markers such as TO, G6P, and cytochrome P450 subtypes and could be infected with HBV. CFPH-chimeric mice with a relatively high replacement rate exhibited viremia and had high serum levels of hepatitis B surface antigen. Conclusion: Serially subcultured human hepatic progenitor-like cells from postnatal livers successfully repopulated injured livers and exhibited several phenotypes of mature hepatocytes, including susceptibility to HBV. In vitro–expanded CFPHs can be used to characterize the differentiation state of human hepatic progenitor-like cells. (HEPATOLOGY 2008.)

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