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Isolation and Fluorescence-Activated Cell Sorting of Mouse Keratinocytes Expressing β-Galactosidase

  • Maria Kasper
  • Rune Toftgård
  • Viljar Jaks
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1453)

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.

Key words

Beta-galactosidase (β-galactosidase) Genetically engineered reporter mice Keratinocytes Skin Epidermis Stem cells FACS sorting 

Notes

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.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Biosciences and Nutrition and Center for Innovative MedicineKarolinska InstitutetHuddingeSweden
  2. 2.Institute of Molecular and Cell BiologyUniversity of TartuTartuEstonia

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