title: Isolation and In Vitro Characterization of Epidermal Stem Cells.
abstract: Colony-forming assays represent prospective methods, where cells isolated from enzymatically dissociated tissues or from tissue cultures are assessed for their proliferative capacity in vitro. Complex tissues such as the epithelial component of the skin (the epidermis) are characterized by a substantial cellular heterogeneity. Analysis of bulk populations of cells by colony-forming assays can consequently be convoluted by a number of factors that are not controlled for in population wide studies. It is therefore advantageous to refine in vitro growth assays by sub-fractionation of cells using flow cytometry. Using markers that define the spatial origin of epidermal cells, it is possible to interrogate the specific characteristics of subpopulations of cells based on their in vivo credentials. Here, we describe how to isolate, culture, and characterize keratinocytes from murine back and tail skin sorted by surface antigens associated with adult stem cell characteristics.
authors.affiliation: Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen N, Denmark.
authors.forename: Kasper S
authors.initials: KS
authors.lastname: Moestrup
authors.affiliation: Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen N, Denmark.
authors.forename: Marianne S
authors.initials: MS
authors.lastname: Andersen
authors.affiliation: Biotech Research and Innovation Centre, Faculty of Health and Medical Sciences, University of Copenhagen, Ole Maaloes Vej 5, 2200, Copenhagen N, Denmark. kim.jensen@bric.ku.dk.
authors.forename: Kim B
authors.initials: KB
authors.lastname: Jensen
mesh_terms: D000818:Animals; D015415:Biomarkers; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D018920:Coculture Techniques; D003114:Colony-Forming Units Assay; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D016130:Immunophenotyping; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D010641:Phenotype; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: D015415:Biomarkers
keywords: Flow cytometry; Isolation; Keratinocyte; Pilosebaceous unit; Skin; Stem cell