Abstract
The skin, one of the largest organs of the body, is a dynamic tissue in which terminally differentiated keratinocytes are replaced by the proliferation and differentiation of epidermal stem cells. Epidermal stem cells are relatively undifferentiated, retain a high capacity for self-renewal throughout their lifetime, and normally have a slow cell division cycle in vivo. Furthermore, they have a high proliferation potential in vitro, and it is often desirable to isolate and culture them from adult mice to use in conjunction with in vivo studies. However, the isolation of these cells has been problematic. Here, we describe reliable methods for identifying a population of isolated bulge stem cells by flow cytometry and for measuring the growth and differentiation potential of primary mouse keratinocytes by clonal analysis.
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References
Terskikh VV, Vasiliev AV, Vorotelyak EA (2012) Label retaining cells and cutaneous stem cells. Stem Cell Rev 8(2):414–425
Blanpain C, Fuchs E (2006) Epidermal stem cells of the skin. Annu Rev Cell Dev Biol 22:339–373
Blanpain C, Lowry WE, Geoghegan A et al (2004) Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche. Cell 118:635–648
Horsley V, Aliprantis AO, Polak L et al (2008) NFATc1 balances quiescence and proliferation of skin stem cells. Cell 132:299–310
Rhee H, Polak L, Fuchs E (2006) Lhx2 maintains stem cell character in hair follicles. Science 312:1946–1949
Watt FM, Jensen KB (2009) Epidermal stem cell diversity and quiescence. EMBO Mol Med 1:260–267
Inoue K, Aoi N, Sato T et al (2009) Differential expression of stem-cell-associated markers in human hair follicle epithelial cells. Lab Invest 89:844–856
Medina RJ, Kataoka K, Takaishi M et al (2006) Isolation of epithelial stem cells from dermis by a three-dimensional culture system. J Cell Biochem 98:174–184
Nowak JA, Fuchs E (2009) Isolation and culture of epithelial stem cells. Methods Mol Biol 482:215–232
Brownell I, Guevara E, Bai CB et al (2011) Nerve-derived sonic hedgehog defines a niche for hair follicle stem cells capable of becoming epidermal stem cells. Cell Stem Cell 8:552–565
Strachan LR, Ghadially R (2008) Tiers of clonal organization in the epidermis: the epidermal proliferation unit revisited. Stem Cell Rev 4:149–157
Greco V, Chen T, Rendl M et al (2009) A two-step mechanism for stem cell activation during hair regeneration. Cell Stem Cell 4:155–169
Barrandon Y, Green H (1987) Three clonal types of keratinocyte with different capacities for multiplication. Proc Natl Acad Sci USA 84:2302–2306
Pellegrini G, Ranno R, Stracuzzi G et al (1999) The control of epidermal stem cells (holoclones) in the treatment of massive full-thickness burns with autologous keratinocytes cultured on fibrin. Transplantation 68:868–879
Yang L, Mao C, Teng Y et al (2005) Targeted disruption of Smad4 in mouse epidermis results in failure of hair follicle cycling and formation of skin tumors. Cancer Res 65:8671–8678
Yang L, Wang L, Yang X (2009) Disruption of Smad4 in mouse epidermis leads to depletion of follicle stem cells. Mol Biol Cell 20:882–890
Acknowledgments
This work was supported by grant 2012CB945100, 2011CB504202 from Chinese National Key Program on Basic Research, grant 31030040, 30871396, and 81123001 from National Natural Science Foundation of China, and 2012ZX10004502 from Chinese Key Project for the Infectious Diseases.
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Wang, Y., Yang, X. (2013). Identification and Analysis of Epidermal Stem Cells from Primary Mouse Keratinocytes. In: Turksen, K. (eds) Skin Stem Cells. Methods in Molecular Biology, vol 989. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-330-5_7
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DOI: https://doi.org/10.1007/978-1-62703-330-5_7
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-329-9
Online ISBN: 978-1-62703-330-5
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