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Isolation and Characterization of Colony-Forming Progenitor Cells from Adult Pancreas

  • Janine C. QuijanoEmail author
  • Jacob R. Tremblay
  • Jeffrey Rawson
  • Hsun Teresa Ku
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2029)

Abstract

Obtaining, growing, and analysis of pancreatic progenitor cells.

Adult stem and progenitor cells have been successfully used for cell-based therapies such as transplantation of hematopoietic stem cells for various diseases. Whether stem and progenitor cells in the adult pancreas can be identified and used for replacement therapy has been a highly controversial topic. To address this controversy, our laboratory has developed in vitro colony assays to detect and characterize individual pancreatic stem and progenitor-like cells. We found that a subpopulation of ductal cells in the adult murine pancreas has the abilities to self-renew and differentiate into multiple pancreatic lineages in three-dimensional space in methylcellulose-containing semisolid media. This protocol details the techniques used for culturing and characterizing these pancreatic stem and progenitor-like cells, which we have named pancreatic colony-forming units (PCFUs), as well as their progenies (colonies). The techniques presented here include dissociation of pancreases, sorting antibody-stained cells with a fluorescence-activated cell sorter, viral transduction of dissociated pancreatic cells, growth of PCFUs in semi-solid media, whole-mount immunostaining and Western blot analysis for proteins expressed in colonies, and kidney capsule transplantation of colonies for in vivo functional analysis.

Key words

Pancreas Progenitor Stem cells Organoid Pancreatic colony-forming units Differentiation Self-renewal 

Notes

Acknowledgments

J.C.Q. is supported by a Juvenile Diabetes Research Foundation (JDRF) Postdoctoral Fellowship 3-PDF-2016-174-A-N. This work is funded in part by a National Institutes of Health (NIH) Grant R01DK099734 to H.T.K.

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Copyright information

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

Authors and Affiliations

  • Janine C. Quijano
    • 1
    Email author
  • Jacob R. Tremblay
    • 1
  • Jeffrey Rawson
    • 1
  • Hsun Teresa Ku
    • 1
    • 2
  1. 1.Department of Translational Research and Cellular Therapeutics, Diabetes and Metabolism Research InstituteBeckman Research Institute of City of HopeDuarteUSA
  2. 2.Irell & Manella Graduate School of Biological SciencesDuarteUSA

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