Functional Characterization of Melanocyte Stem Cells in Hair Follicles

In mice, coat pigmentation requires a stem cell (SC) system in which the survival, proliferation, and differentiation of melanocytes (MCs) are regulated by microenvironments in hair follicles (HFs). In vitro systems are required to analyze the behavior of single melanocyte stem cells (MCSCs) and the...

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Bibliographic Details
Published in:Journal of investigative dermatology 2011-12, Vol.131 (12), p.2358-2367
Main Authors: Nishikawa-Torikai, Satomi, Osawa, Masatake, Nishikawa, Shin-ichi
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - pharmacology
Biological and medical sciences
Cell Differentiation - drug effects
Cells, Cultured
Dermatology
Female
Fibroblast Growth Factor 2 - metabolism
Fibroblast Growth Factor 2 - pharmacology
Hair Follicle - drug effects
Hair Follicle - metabolism
Medical sciences
Melanocytes - drug effects
Melanocytes - metabolism
Mice
Proto-Oncogene Proteins c-kit - antagonists & inhibitors
Proto-Oncogene Proteins c-kit - metabolism
Stem Cell Factor - pharmacology
Stem Cells - drug effects
Stem Cells - metabolism
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Summary:In mice, coat pigmentation requires a stem cell (SC) system in which the survival, proliferation, and differentiation of melanocytes (MCs) are regulated by microenvironments in hair follicles (HFs). In vitro systems are required to analyze the behavior of single melanocyte stem cells (MCSCs) and their potential to form SC systems in vivo. We describe here an experimental system for the isolation, self-renewal, and differentiation of MCSCs, as well as an in vivo reconstitution assay for assessing their potential. Using Dcttm1(Cre)Bee/CAG-CAT-GFP mice, we show that, in the presence of stem cell factor and basic fibroblast growth factor and the XB2 feeder cell line, purified MCSCs can undergo clonogenic proliferation, resulting in c-Kitlow side scatterlow cells. In culture, these cells maintain their capacity to differentiate and reconstitute an MCSC system in HFs. As these cells are present in the upper part of the HF near the bulge region, express only low levels of housekeeping genes, and are resistant to neonatal treatment with ACK2, it is likely that only MCSCs that are quiescent in vivo have clonogenic activity in vitro. We also found that MCSCs can be purified from wild-type mice by fluorescent cell sorting and can be characterized in vitro.
ISSN:0022-202X
1523-1747
DOI:10.1038/jid.2011.195