Clonal culture of adult mouse lung epithelial stem/progenitor cells.
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.
Lung Health Research Centre, Department of Pharmacology and Therapeutics, University of Melbourne, Grattan Street, Parkville, VIC, 3010, Australia, jlmcq@unimelb.edu.au.
Jonathan L
JL
McQualter
Ivan
I
Bertoncello
D053687:Adult Stem Cells / Q000166:cytology*; D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis; D051927:CD24 Antigen / Q000032:analysis; D015815:Cell Adhesion Molecules / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay / Q000379:methods; D000071858:Epithelial Cell Adhesion Molecule; D004847:Epithelial Cells / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods*; D039663:Integrin beta4 / Q000032:analysis; D017493:Leukocyte Common Antigens / Q000032:analysis; D008168:Lung / Q000166:cytology*; D051379:Mice; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis
D000951:Antigens, Neoplasm; D051927:CD24 Antigen; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule; D039663:Integrin beta4; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D017493:Leukocyte Common Antigens