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Keratinocyte Differentiation by Flow Cytometry

  • Natalia Sanz-Gómez
  • Ana Freije
  • Alberto GandarillasEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2109)

Abstract

The epidermis is continuously exposed to environmental hazard and undergoes continuous cell renewal. The maintenance of the epidermal balance between proliferation and differentiation is essential for the homeostasis of the skin. Proliferation and terminal differentiation are compartmentalized in basal and suprabasal layers, respectively. These compartments can be identified by different patterns of protein expression that can be used as differentiation markers. For instance, components of the intermediate filament cytoskeleton keratins K5 and K14 are confined to the proliferative basal layer, while keratins K1 and K10, keratins K6 and K16, or precursors of the cornified envelope such as involucrin are expressed by suprabasal terminally differentiating cells. The analysis of the expression of these markers allows studying the imbalance typical of disease. Although these markers have been traditionally analyzed on skin microsections, on attached cells by immunostaining or by western blotting, it is possible and advantageous to quantify them by flow cytometry. We have extensively applied this technology onto human and mouse keratinocytes. Here we describe detailed flow cytometry methods to determine the differentiation status of keratinocyte populations.

Keywords

Epidermis Differentiation Flow cytometry Keratin K1 Keratin K10 Keratin K13  Keratin K16 Involucrin 

Notes

Acknowledgments

This work was funded by Instituto de Salud Carlos III/FEDER (AG; Spain), grants PI14/00900 and PI17/01307. NSG is recipient of a predoctoral scholarship from Universidad de Cantabria/IDIVAL (Spain). We thank Dr. Fiona Watt for providing SY3 antibody.

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Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Natalia Sanz-Gómez
    • 1
  • Ana Freije
    • 1
  • Alberto Gandarillas
    • 1
    • 2
    Email author
  1. 1.Cell Cycle, Stem Cell Fate and Cancer LaboratoryInstitute for Research Marqués de Valdecilla (IDIVAL)SantanderSpain
  2. 2.INSERM, Languedoc-RoussillonMontpellierFrance

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