Advertisement

Identification and Isolation of Clonogenic Cholangiocyte in Mouse

  • Bin Li
  • Craig Dorrell
  • Pamela S. Canady
  • Leslie Wakefield
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1905)

Abstract

Cholangiocytes are proliferative and are one of the sources for liver progenitor cells. Clonogenic cholangiocytes are defined as cells capable of clonally proliferating and differentiating cholangiocytes both in vitro and in vivo. In this protocol, we describe the method for isolation of primary cholangiocytes from mouse. To study the heterogeneity of cholangiocytes, we used flow cytometry-based cell sorting to isolate different subsets of cholangiocytes. Organoid-forming efficiencies from sorted single cells are compared within different cholangiocyte populations to identify clonogenic cholangiocytes.

Key words

Duct cells ST14 FACS Organoid 

Notes

Acknowledgments

The authors thank members in the Markus Grompe Lab at Oregon Health and Science University for technical supports. The authors also thank Oregon Health and Science University Flow Cytometry Core for technical assistance.

References

  1. 1.
    Dorrell C et al (2011) Prospective isolation of a bipotential clonogenic liver progenitor cell in adult mice. Genes Dev 25:1193–1203.  https://doi.org/10.1101/gad.2029411CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Azuma H et al (2007) Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice. Nat Biotechnol 25:903–910.  https://doi.org/10.1038/nbt1326CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Li B et al (2017) Adult mouse liver contains two distinct populations of cholangiocytes. Stem Cell Reports 9:478–489.  https://doi.org/10.1016/j.stemcr.2017.06.003CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Huch M et al (2013) In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature 494:247–250.  https://doi.org/10.1038/nature11826CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Dorrell C et al (2014) The organoid-initiating cells in mouse pancreas and liver are phenotypically and functionally similar. Stem Cell Res 13:275–283.  https://doi.org/10.1016/j.scr.2014.07.006CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Bin Li
    • 1
  • Craig Dorrell
    • 1
  • Pamela S. Canady
    • 1
  • Leslie Wakefield
    • 1
  1. 1.Oregon Stem Cell Center, Oregon Health and Science UniversityPortlandUSA

Personalised recommendations