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Detection, Labeling, and Culture of Lung Stem and Progenitor Cells

  • Ivan Bertoncello
  • Gianni Carraro
  • Jonathan L. McQualterEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1842)

Abstract

Identification, isolation, and clonal culture of stem cells is essential for understanding their proliferative and differentiation potential, and the cellular and molecular mechanisms that regulate their fate. Akin to development in vivo, the in vitro growth of adult lung epithelial stem cells requires support of mesenchymal-derived growth factors. In the adult mouse lung, epithelial stem/progenitor cells are defined by the phenotype CD45neg CD31neg EpCAMpos CD104pos CD24low, and mesenchymal cells are defined by the phenotype CD45neg CD31neg EpCAMneg Sca-1hi. Here we describe a method for primary cell isolation from the adult mouse lung, a flow cytometry strategy for fractionation of epithelial stem/progenitor cells and mesenchymal cells, and a three-dimensional epithelial colony-forming assay.

Key words

Lung epithelium Stem cells Colony-forming assay Flow cytometry 

Notes

Acknowledgments

This work was supported by grants from RMIT University and the Australian National Health and Medical Research Council (NHMRC).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Ivan Bertoncello
    • 1
  • Gianni Carraro
    • 2
  • Jonathan L. McQualter
    • 3
    Email author
  1. 1.Lung Health Research Centre, Department of Pharmacology and TherapeuticsUniversity of MelbourneParkvilleAustralia
  2. 2.Lung and Regenerative Medicine Institutes, Department of MedicineCedars-Sinai Medical CenterLos AngelesUSA
  3. 3.School of Health and Biomedical SciencesRMIT UniversityBundooraAustralia

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