Abstract
In this article, we provide a method to isolate primary epidermal melanocytes from reporter mice, which also allow targeted gene inactivation. The mice from which these cells are isolated are bred into a Rosa26mT/mG reporter background, which results in GFP expression in the targeted melanocytic cell population. These cells are isolated and cultured to >95% purity. The cells can be used for gene expression studies, clonogenic experiments, and biological assays, such as capacity for migration. Melanocytes are slow moving cells, and we also provide a method to measure motility using individual cell tracking and data analysis.
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Acknowledgments
This work was supported by grants to LD from the Canadian Institutes of Health Research, the National Science and Engineering Research Council, and the Cancer Research Society. MC is the recipient of an Ontario Graduate Scholarship from the Ontario Ministry of Advanced Education and Skills Development.
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Dagnino, L., Crawford, M. (2018). Isolation, Culture, and Motility Measurements of Epidermal Melanocytes from GFP-Expressing Reporter Mice. In: Turksen, K. (eds) Skin Stem Cells. Methods in Molecular Biology, vol 1879. Humana Press, New York, NY. https://doi.org/10.1007/7651_2018_116
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DOI: https://doi.org/10.1007/7651_2018_116
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Publisher Name: Humana Press, New York, NY
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