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Skin Stem Cells pp 101–110Cite as

Interfollicular Epidermal Stem Cells: Boosting and Rescuing from Adult Skin

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1879))

Abstract

Epidermal stem cells (EpSCs) isolation struggle remains, mainly due to the yet essential requirement of well-defined approaches and markers. The herein proposed methodology integrates an assemblage of strategies to accomplish the enrichment of the interfollicular EpSCs multipotent fraction and their subsequent separation from the remaining primary human keratinocytes (hKC) culture. Those include rapid adherence of freshly isolated hKC to collagen type IV through the β1-integrin ligand and Rho-associated protein kinase inhibitor (Rocki) Y-27632 administration to the cultures, followed by an immunomagnetic separation to obtain populations based in the combined CD49fbri/CD71dim expression. Flow cytometry is the supporting method to analyze the effect of the treatments over the expression rate of early epidermal markers keratins19/5/14 and in correlation to CD49fbri/CD71dim sub-populations. The step-by-step methodology herein described indulges the boosting and consecutive purification and separation of interfollicular epidermal stem cells, from human keratinocytes cultures.

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Author information

Authors and Affiliations

  1. 3B’s Research Group—Biomaterials, Biodegradable and Biomimetics, Avepark—Parque de Ciência e Tecnologia, Zona Industrial da Gandra, Guimarães, Portugal

    Mariana T. Cerqueira, Ana M. Frias, Rui L. Reis & Alexandra P. Marques

  2. ICVS/3B’s—PT Government Associate Laboratory, Braga/Guimarães, Portugal

    Mariana T. Cerqueira, Ana M. Frias, Rui L. Reis & Alexandra P. Marques

  3. The Discoveries Centre for Regenerative and Precision Medicine, Headquarters at University of Minho, Guimarães, Portugal

    Rui L. Reis & Alexandra P. Marques

Authors
  1. Mariana T. Cerqueira
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  2. Ana M. Frias
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  3. Rui L. Reis
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  4. Alexandra P. Marques
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Corresponding author

Correspondence to Alexandra P. Marques .

Editor information

Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

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Cerqueira, M.T., Frias, A.M., Reis, R.L., Marques, A.P. (2018). Interfollicular Epidermal Stem Cells: Boosting and Rescuing from Adult Skin. In: Turksen, K. (eds) Skin Stem Cells. Methods in Molecular Biology, vol 1879. Humana Press, New York, NY. https://doi.org/10.1007/7651_2018_146

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  • DOI: https://doi.org/10.1007/7651_2018_146

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8869-3

  • Online ISBN: 978-1-4939-8870-9

  • eBook Packages: Springer Protocols

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