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Isolation, Culture, and Motility Measurements of Epidermal Melanocytes from GFP-Expressing Reporter Mice

  • Lina DagninoEmail author
  • Melissa Crawford
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1879)

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.

Keywords

Cell migration Cre recombinase GFP Melanocyte 

Notes

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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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  1. 1.Department of Physiology and Pharmacology, Child Health Research Institute and Lawson Health Research InstituteUniversity of Western OntarioLondonCanada
  2. 2.Department of Physiology and PharmacologyUniversity of Western OntarioLondonCanada

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