Isolation of Bipotential Liver Progenitor Cells from Neonatal Mouse Liver

  • Naoki TanimizuEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1905)


Liver stem/progenitor cells (LPCs) are defined as bipotential cells differentiating into both hepatocytes and cholangiocytes. The Notch, TGFβ, and Hippo pathways have been implicated in lineage determination of LPCs during development and regeneration. However, the molecular mechanisms governing the lineage specification have not been fully elucidated, yet. Epithelial adhesion molecule (EpCAM) is a marker of cholangiocytes and of LPCs. We found that EpCAM+ cells isolated from neonatal liver contain LPCs that clonally proliferate and are bipotential in vitro and in vivo. Furthermore, EpCAM+ progenies keep the capacity of bidirectional differentiation even after long-term culture. These cells are useful to investigate the molecular mechanisms regulating lineage commitment and epithelial differentiation of LPCs.

Key words

Tissue stem cell Liver Neonate Bipotential Hepatocyte Cholangiocyte Liver stem/progenitor cell Fluorescence-activated cell sorting Colony assay Clonal culture 



I thank Dr. Toshihiro Mitaka and Dr. Norihisa Ichinohe for helpful discussion and Ms. Yumiko Tsukamoto and Ms. Minako Kuwano for technical assistance. This work is supported by the Ministry of Education, Culture, Sports, Science and Technology, Japan, Grants-in-Aid for Scientific Research (C) (25460271, 16 K08716), and Grants-in-Aid for Scientific Research on Innovative Areas “Stem Cell Aging and Disease” (17H05653).


  1. 1.
    Miyajima A, Tanaka M, Itoh T (2014) Stem Progenitor cell sin liver development, homeostasis, regeneration, and reprogramming. Cell Stem Cell 14:261–274CrossRefGoogle Scholar
  2. 2.
    Espanol-Suner R, Carpentier R, Van HN et al (2012) Liver progenitor cells yield functional hepatocytes in response to chronic liver injury in mice. Gastroenterology 143:1564–1575 e1567CrossRefGoogle Scholar
  3. 3.
    Malato Y, Naqvi S, Schurmann N et al (2011) Fate tracing of mature hepatocytes in mouse liver homeostasis and regeneration. J Clin Invest 121:4850–4860CrossRefGoogle Scholar
  4. 4.
    Choi TY, Ninov N, Stainer DY et al (2014) Extensive conversion of hepatic biliary epithelial cells to hepatocytes after near total loss of hepatocytes in zebrafish. Gastroenterology 146:776–788CrossRefGoogle Scholar
  5. 5.
    Raven A, Lu WY, Man TY et al (2017) Cholangiocytes act as facultative liver stem cells during impaired hepatocyte regeneration. Nature 547:350–354CrossRefGoogle Scholar
  6. 6.
    Kamimoto K, Kaneko K, Kok CY et al (2016) Heterogeneity and stochastic growth regulation of biliary epithelial cells dictate dynamic epithelial tissue remodeling. Elife 5:e15034Google Scholar
  7. 7.
    Li B, DorrellC CPS et al (2017) Adult mouse liver contains two distinct populations of cholangiocytes. Stem Cell Reports 9:478–489CrossRefGoogle Scholar
  8. 8.
    Tanimizu N, Kobayashi S, Ichinohe N et al (2014) Downregulation of miR122 by grainyhead-like 2 restricts the hepatocytic differentiation potential of adult liver progenitor cells. Development 141:4448–4456CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Tissue Development and RegenerationResearch Institute for Frontier Medicine, Sapporo Medical University School of MedicineSapporo, HokkaidoJapan

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