Clonal culture of adult mouse lung epithelial stem/progenitor cells

Methods Mol Biol. 2015:1235:231-41. doi: 10.1007/978-1-4939-1785-3_17.

Abstract

Clonal culture of stem cells is crucial for their identification, and the characterization of the cellular and molecular mechanisms that regulate their proliferation and differentiation. In the adult mouse lung, epithelial stem/progenitor cells are defined by the phenotype CD45(neg) CD31(neg) EpCAM(pos) CD104(pos) CD24(low). Here we describe a tissue dissociation and flow cytometry strategy for the detection and isolation of adult mouse lung epithelial stem/progenitor cells, and a three-dimensional colony-forming assay for their clonal culture in vitro.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells / cytology*
  • Animals
  • Antigens, Neoplasm / analysis
  • CD24 Antigen / analysis
  • Cell Adhesion Molecules / analysis
  • Cell Culture Techniques / methods
  • Cell Proliferation
  • Cell Separation / methods*
  • Cells, Cultured
  • Colony-Forming Units Assay / methods
  • Epithelial Cell Adhesion Molecule
  • Epithelial Cells / cytology*
  • Flow Cytometry / methods*
  • Integrin beta4 / analysis
  • Leukocyte Common Antigens / analysis
  • Lung / cytology*
  • Mice
  • Platelet Endothelial Cell Adhesion Molecule-1 / analysis

Substances

  • Antigens, Neoplasm
  • CD24 Antigen
  • Cell Adhesion Molecules
  • Epithelial Cell Adhesion Molecule
  • Integrin beta4
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Leukocyte Common Antigens