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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media New York
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/7651_2018_152
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