Characterizing the phenotype of murine epidermal progenitor cells: complementary whole-mount visualization and flow cytometry strategies.
The epidermis and its appendages, the hair follicle and sebaceous gland, have the capacity to constantly regenerate throughout adult life. Postnatal hair follicles undergo a cyclic mode of tissue homeostasis, defined by periods of growth, degeneration, and rest. A multipotent population of stem cells residing within the hair follicle bulge not only generates the hair lineages during each hair cycle, but also transiently contributes to the repair of epidermis following wounding. In this chapter, we provide methods for identifying epidermal stem cells and investigating their proliferative and apoptotic characteristics. We introduce whole-mount and flow cytometry techniques, which complement each other by permitting visualization of the epidermal stem cell compartment in situ and assessment of the phenotype of purified cells. These techniques can easily be adapted to characterize novel putative epidermal stem or progenitor cell populations. By applying whole-mount and flow cytometry techniques to characterize normal and genetically modified mice with skin defects, we expect to learn more about the factors that regulate stem cell self-renewal and differentiation.
Centre for Cutaneous Research, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, London, UK.
Korinna D
KD
Henseleit
Ann P
AP
Wheeler
Gary
G
Warnes
Kristin M
KM
Braun
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D002453:Cell Cycle; D000078404:Epidermal Cells; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D051379:Mice; D018613:Microscopy, Confocal; D057054:Molecular Imaging / Q000379:methods*; D010641:Phenotype; D013194:Staining and Labeling; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D017304:Annexin A5