title: Isolation and Fluorescence-Activated Cell Sorting of Mouse Keratinocytes Expressing β-Galactosidase.
abstract: During the past decade, the rapid development of new transgenic and knock-in mouse models has propelled epidermal stem-cell research into "fast-forward mode". It has become possible to identify and visualize defined cell populations during normal tissue maintenance, and to follow their progeny during the processes of homeostasis, wound repair, and tumorigenesis. Moreover, these cells can be isolated using specific labels, and characterized in detail using an array of molecular and cell biology approaches. The bacterial enzyme, β-galactosidase (β-gal), the product of the LacZ gene, is one of the most commonly used in vivo cell labels in genetically-engineered mice. The protocol described in this chapter provides a guideline for the isolation of viable murine epidermal cells expressing β-gal, which can then be subjected to further characterization in vivo or in vitro.
authors.affiliation: Department of Biosciences and Nutrition and Center for Innovative Medicine, Karolinska Institutet, Novum, 141 83, Huddinge, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Kasper
authors.affiliation: Department of Biosciences and Nutrition and Center for Innovative Medicine, Karolinska Institutet, Novum, 141 83, Huddinge, Sweden.
authors.forename: Rune
authors.initials: R
authors.lastname: Toftgård
authors.affiliation: Department of Biosciences and Nutrition and Center for Innovative Medicine, Karolinska Institutet, Novum, 141 83, Huddinge, Sweden. viljar.jaks@ut.ee.|Institute of Molecular and Cell Biology, University of Tartu, Riia 23, 51010, Tartu, Estonia. viljar.jaks@ut.ee.
authors.forename: Viljar
authors.initials: V
authors.lastname: Jaks
mesh_terms: D000818:Animals; D000078404:Epidermal Cells; D005434:Flow Cytometry / Q000379:methods; D015870:Gene Expression; D017930:Genes, Reporter; D015603:Keratinocytes / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001616:beta-Galactosidase
keywords: Beta-galactosidase (β-galactosidase); Epidermis; FACS sorting; Genetically engineered reporter mice; Keratinocytes; Skin; Stem cells