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Isolation of Human Skin Epidermal Stem Cells Based on the Expression of Endothelial Protein C Receptor

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Skin Stem Cells

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

Abstract

Skin epidermis is a continuous self-renewal tissue maintained by interfollicular epidermal stem cells (IESCs) that reside in the basal layer of epidermis. IESCs also contribute to the repair and regeneration of the epidermis during wound healing. The great plasticity and easy accessibility afforded by IESCs make them a promising source of stem cells for scientific research and clinical applications. Thus, simple methods to isolate and define pure and viable IESCs are a valuable resource. Here, we provide a method for isolating IESCs from human skin epidermis. This method relies exclusively on selecting cells with a higher expression of the endothelial protein C receptor, using fluorescence-activated cell sorting.

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References

  1. Fuchs E, Raghavan S (2002) Getting under the skin of epidermal morphogenesis. Nat Rev Genet 3(3):199–209. https://doi.org/10.1038/nrg758

    Article  CAS  PubMed  Google Scholar 

  2. Blanpain C, Fuchs E (2014) Stem cell plasticity. Plasticity of epithelial stem cells in tissue regeneration. Science 344(6189):1242281. https://doi.org/10.1126/science.1242281

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Hirsch T, Rothoeft T, Teig N, Bauer JW, Pellegrini G, De Rosa L, Scaglione D, Reichelt J, Klausegger A, Kneisz D, Romano O, Secone Seconetti A, Contin R, Enzo E, Jurman I, Carulli S, Jacobsen F, Luecke T, Lehnhardt M, Fischer M, Kueckelhaus M, Quaglino D, Morgante M, Bicciato S, Bondanza S, De Luca M (2017) Regeneration of the entire human epidermis using transgenic stem cells. Nature 551(7680):327–332. https://doi.org/10.1038/nature24487

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Jackson CJ, Tonseth KA, Utheim TP (2017) Cultured epidermal stem cells in regenerative medicine. Stem Cell Res Ther 8(1):155. https://doi.org/10.1186/s13287-017-0587-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Jones PH, Watt FM (1993) Separation of human epidermal stem cells from transit amplifying cells on the basis of differences in integrin function and expression. Cell 73(4):713–724

    Article  CAS  Google Scholar 

  6. Zhou JX, Chen SY, Liu WM, Cao YJ, Duan EK (2004) Enrichment and identification of human ‘fetal’ epidermal stem cells. Hum Reprod 19(4):968–974. https://doi.org/10.1093/humrep/deh166

    Article  PubMed  Google Scholar 

  7. Papini S, Cecchetti D, Campani D, Fitzgerald W, Grivel JC, Chen S, Margolis L, Revoltella RP (2003) Isolation and clonal analysis of human epidermal keratinocyte stem cells in long-term culture. Stem Cells 21(4):481–494. https://doi.org/10.1634/stemcells.21-4-481

    Article  PubMed  Google Scholar 

  8. Li A, Simmons PJ, Kaur P (1998) Identification and isolation of candidate human keratinocyte stem cells based on cell surface phenotype. Proc Natl Acad Sci U S A 95(7):3902–3907

    Article  CAS  Google Scholar 

  9. Kaur P, Li A (2000) Adhesive properties of human basal epidermal cells: an analysis of keratinocyte stem cells, transit amplifying cells, and postmitotic differentiating cells. J Invest Dermatol 114(3):413–420. https://doi.org/10.1046/j.1523-1747.2000.00884.x

    Article  CAS  PubMed  Google Scholar 

  10. Li J, Miao C, Guo W, Jia L, Zhou J, Ma B, Peng S, Liu S, Cao Y, Duan E (2008) Enrichment of putative human epidermal stem cells based on cell size and collagen type IV adhesiveness. Cell Res 18(3):360–371. https://doi.org/10.1038/cr.2007.103

    Article  CAS  PubMed  Google Scholar 

  11. Dunnwald M, Tomanek-Chalkley A, Alexandrunas D, Fishbaugh J, Bickenbach JR (2001) Isolating a pure population of epidermal stem cells for use in tissue engineering. Exp Dermatol 10(1):45–54

    Article  CAS  Google Scholar 

  12. Xue M, Dervish S, Chan B, Jackson CJ (2017) The endothelial protein C receptor is a potential stem cell marker for epidermal keratinocytes. Stem Cells 35(7):1786–1798. https://doi.org/10.1002/stem.2630

    Article  CAS  PubMed  Google Scholar 

  13. Balazs AB, Fabian AJ, Esmon CT, Mulligan RC (2006) Endothelial protein C receptor (CD201) explicitly identifies hematopoietic stem cells in murine bone marrow. Blood 107(6):2317–2321

    Article  CAS  Google Scholar 

  14. Matsumura W, Fujita Y, Nakayama C, Shinkuma S, Suzuki S, Nomura T, Abe R, Shimizu H (2018) Establishment of integration-free induced pluripotent stem cells from human recessive dystrophic epidermolysis bullosa keratinocytes. J Dermatol Sci 89(3):263–271. https://doi.org/10.1016/j.jdermsci.2017.11.017

    Article  CAS  PubMed  Google Scholar 

  15. Mavilio F, Pellegrini G, Ferrari S, Di Nunzio F, Di Iorio E, Recchia A, Maruggi G, Ferrari G, Provasi E, Bonini C, Capurro S, Conti A, Magnoni C, Giannetti A, De Luca M (2006) Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells. Nat Med 12(12):1397–1402. https://doi.org/10.1038/nm1504

    Article  CAS  PubMed  Google Scholar 

  16. Kratz G, Haegerstrand A, Dalsgaard CJ (1991) Conditioned medium from cultured human keratinocytes has growth stimulatory properties on different human cell types. J Invest Dermatol 97(6):1039–1043

    Article  CAS  Google Scholar 

  17. Wu Y, Wu J, Lee DY, Yee A, Cao L, Zhang Y, Kiani C, Yang BB (2005) Versican protects cells from oxidative stress-induced apoptosis. Matrix Biol 24(1):3–13. https://doi.org/10.1016/j.matbio.2004.11.007

    Article  CAS  PubMed  Google Scholar 

  18. Hirai H, Umegaki R, Kino-Oka M, Taya M (2002) Characterization of cellular motions through direct observation of individual cells at early stage in anchorage-dependent culture. J Biosci Bioeng 94(4):351–356

    Article  CAS  Google Scholar 

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Correspondence to Meilang Xue .

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Xue, M., Dervish, S., Jackson, C.J. (2018). Isolation of Human Skin Epidermal Stem Cells Based on the Expression of Endothelial Protein C Receptor. 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_152

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  • DOI: https://doi.org/10.1007/7651_2018_152

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  • 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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