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.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Mansour SL, Thomas KR, Deng CX et al (1990) Introduction of a lacZ reporter gene into the mouse int-2 locus by homologous recombination. Proc Natl Acad Sci U S A 87:7688–7692
Morrey JD, Bourn SM, Bunch TD et al (1991) In vivo activation of human immunodeficiency virus type 1 long terminal repeat by UV type A (UV-A) light plus psoralen and UV-B light in the skin of transgenic mice. J Virol 65:5045–5051
Ikawa M, Kominami K, Yoshimura Y et al (1995) Green fluorescent protein as a marker in transgenic mice. Dev Growth Differ 37:455–459
Snippert HJ, van der Flier LG, Sato T et al (2010) Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143:134–144
Bickenbach JR, Chism E (1998) Selection and extended growth of murine epidermal stem cells in culture. Exp Cell Res 244:184–195
Trempus CS, Morris RJ, Bortner CD et al (2003) Enrichment for living murine keratinocytes from the hair follicle bulge with the cell surface marker CD34. J Invest Dermatol 120:501–511
Tani H, Morris RJ, Kaur P (2000) Enrichment for murine keratinocyte stem cells based on cell surface phenotype. Proc Natl Acad Sci U S A 97:10960–10965
Jensen KB, Collins CA, Nascimento E et al (2009) Lrig1 expression defines a distinct multipotent stem cell population in mammalian epidermis. Cell Stem Cell 4:427–439
Nijhof JG, Braun KM, Giangreco A et al (2006) The cell-surface marker MTS24 identifies a novel population of follicular keratinocytes with characteristics of progenitor cells. Development 133:3027–3037
Tumbar T, Guasch G, Greco V et al (2004) Defining the epithelial stem cell niche in skin. Science 303:359–363
Takeda N, Jain R, Leboeuf MR et al (2013) Hopx expression defines a subset of multipotent hair follicle stem cells and a progenitor population primed to give rise to K6+ niche cells. Development 140:1655–1664
Snippert HJ, Haegebarth A, Kasper M et al (2010) Lgr6 marks stem cells in the hair follicle that generate all cell lineages of the skin. Science 327:1385–1389
Barker N, van Es JH, Kuipers J et al (2007) Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449:1003–1007
Jaks V, Barker N, Kasper M et al (2008) Lgr5 marks cycling, yet long-lived, hair follicle stem cells. Nat Genet 40:1291–1299
Wu WY, Morris RJ (2005) Method for the harvest and assay of in vitro clonogenic keratinocytes stem cells from mice. Methods Mol Biol 289:79–86
Philpott MP, Green MR, Kealey T (1992) Rat hair follicle growth in vitro. Br J Dermatol 127:600–607
Rendl M, Lewis L, Fuchs E (2005) Molecular dissection of mesenchymal-epithelial interactions in the hair follicle. PLoS Biol 3:e331
Rogers G, Martinet N, Steinert P et al (1987) Cultivation of murine hair follicles as organoids in a collagen matrix. J Invest Dermatol 89:369–379
Acknowledgements
V.J. is supported by an EMBO Installation Grant and two grants, nos. ETF8932 and PUT4 from the Estonian Research Council. M.K. is supported by grants from the Swedish Research Council, the Swedish Cancer Society, the Karolinska Institutet, the Harald and Greta Jeanssons Foundation, the Swedish Foundation for Strategic Research, the Center for Innovative Medicine, and the Ragnar Söderberg Foundation. R.T. is supported by grants from the Swedish Research Council and the Swedish Cancer Society.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kasper, M., Toftgård, R., Jaks, V. (2016). Isolation and Fluorescence-Activated Cell Sorting of Mouse Keratinocytes Expressing β-Galactosidase. In: Hoffman, R. (eds) Multipotent Stem Cells of the Hair Follicle. Methods in Molecular Biology, vol 1453. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3786-8_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3786-8_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3784-4
Online ISBN: 978-1-4939-3786-8
eBook Packages: Springer Protocols