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Isolation and Expansion of Rat Hepatocytic Progenitor Cells

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Hepatic Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1905))

Abstract

This protocol showed procedures to isolate and expand small hepatocytes (SHs), as hepatocytic progenitor cells, from a rat liver. SHs are identified as a subpopulation of mature hepatocytes in a healthy liver. SHs can proliferate to form colonies in serum-free medium on hyaluronic acid-coated dishes, of which cells show CD44 positivity (CD44+ SHs). CD44+ SHs can be separated and purified from colonies by using anti-CD44 antibodies after enzymatic dissociation. CD44+ SHs can proliferate to form colonies on Engelbreth-Holm-Swarm gel (EHS-gel)-coated dishes in the serum-free medium for a long period and subculture for several times. Even after the second passage, the cells possess characteristics of hepatocytes such as expression of albumin and HNF4α. In addition, when the cells are treated with EHS-gel, they can recover highly differentiated functions of hepatocytes such as glycogen production, CYP activity, and bile secretion.

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Author notes

  1. All the authors contributed equally to this work.

Authors and Affiliations

  1. Department of Tissue Development and Regeneration, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan

    Junichi Kino, Norihisa Ichinohe, Masayuki Ishii & Toshihiro Mitaka

  2. Tokushima Research Institute, Otsuka Pharmaceutical Co. Ltd., Tokushima, Japan

    Junichi Kino

  3. Department of Surgery, Surgical Oncology and Science, Sapporo Medical University School of Medicine, Sapporo, Japan

    Masayuki Ishii

Authors
  1. Junichi Kino
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  2. Norihisa Ichinohe
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  3. Masayuki Ishii
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  4. Toshihiro Mitaka
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Corresponding author

Correspondence to Toshihiro Mitaka .

Editor information

Editors and Affiliations

  1. Sapporo Medical University, Sapporo, Hokkaido, Japan

    Naoki Tanimizu

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Kino, J., Ichinohe, N., Ishii, M., Mitaka, T. (2019). Isolation and Expansion of Rat Hepatocytic Progenitor Cells. In: Tanimizu, N. (eds) Hepatic Stem Cells . Methods in Molecular Biology, vol 1905. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8961-4_4

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  • DOI: https://doi.org/10.1007/978-1-4939-8961-4_4

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8960-7

  • Online ISBN: 978-1-4939-8961-4

  • eBook Packages: Springer Protocols

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