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One-Step Simple Isolation Method to Obtain Both Epidermal and Dermal Stem Cells from Human Skin Specimen

  • Hua Qian
  • Xue Leng
  • Jie Wen
  • Qian Zhou
  • Xin Xu
  • Xunwei WuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1879)

Abstract

Stem cells play a crucial role in maintaining and repairing tissues during homeostasis and following injury. The efficient procurement of high quantity and quality of skin stem cells is important for both laboratory studies and clinical applications. Here, we describe a one-step isolation procedure to efficiently obtain both epidermal and dermal cell population from human skin specimen, based on the different influence of the Rho kinase inhibitor Y27632 on the growth of epidermal and dermal cells during the initial culture. Compared with the conventional methods, our protocol shows that it is simpler and less time consuming and can efficiently obtain the high quality of skin stem cells and can maintain the stem cell features after culture expansion.

Keywords

Adult Culture Dermal fibroblasts Epidermal stem cells Human Y-27632 

Notes

Acknowledgement

This work was supported by National Key Research and Development Program of China (2017YFA0104604), General Program of National Natural Science Foundation of China (81772093), and Shandong Taishan Scholar Award (tshw201502065).

References

  1. 1.
    Prodinger CM, Reichelt J, Bauer JW, Laimer M (2017) Current and future perspectives of stem cell therapy in dermatology. Ann Dermatol 29(6):667–687CrossRefGoogle Scholar
  2. 2.
    Gonzales KAU, Fuchs E (2017) Skin and its regenerative powers: an alliance between stem cells and their niche. Dev Cell 43(4):387–401.  https://doi.org/10.1016/j.devcel.10.001CrossRefPubMedGoogle Scholar
  3. 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):327–332CrossRefGoogle Scholar
  4. 4.
    Terunuma A, Limgala RP, Park CJ, Choudhary I, Vogel JC (2010) Efficient procurement of epithelial stem cells from human tissue specimens using a rho-associated protein kinase inhibitor Y-27632. Tissue Eng A 16(4):1363–1368.  https://doi.org/10.1089/ten.TEA.2009.0339CrossRefGoogle Scholar
  5. 5.
    Aasen T, Izpisua Belmonte JC (2010) Isolation and cultivation of human keratinocytes from skin or plucked hair for the generation of induced pluripotent stem cells. Nat Protoc 5(2):371–382.  https://doi.org/10.1038/nprot.2009.241CrossRefPubMedGoogle Scholar
  6. 6.
    Zou D, Pan J, Zhang P, Wu X (2016) A new method to isolate human epidermal keratinocytes. J Clin Dermatol 6:424–429. (in Chinese)Google Scholar
  7. 7.
    Wen J, Zu T, Zhou Q, Leng X, Wu X (2017) Y-27632 simplifies the isolation procedure of human primary epidermal cells by selectively blocking focal adhesion of dermal cells. J Tissue Eng Regen Med.  https://doi.org/10.1002/term.2526CrossRefGoogle Scholar
  8. 8.
    Zhou Q, Zhang Q, Leng X, Zu T, Qin J, Wen J, Wu X (2017) Culture-expanded adult dermal cells derived from scalp tissue are multi- differentiation potential. Smart healthcare, 3(3):1-8 DOI: https://doi.org/10.19335/j.cnki.2096., (in chinese)
  9. 9.
    Watt FM (1998) Epidermal stem cells: markers, patterning and the control of stem cell fate. Philos Trans R Soc Lond B Biol Sci 353(1370):831–837.  https://doi.org/10.1098/rstb.1998.0247CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Barrandon Y, Green H (1987) Three clonal types of keratinocyte with different capacities for multiplication. Proc Natl Acad Sci U S A 84(8):2302–2306CrossRefGoogle Scholar
  11. 11.
    Chen S, Lewallen M, Xie T (2013) Adhesion in the stem cell niche: biological roles and regulation. Development 140(2):255–265.  https://doi.org/10.1242/dev.083139CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Bartsch G, Yoo JJ, De Coppi P, Siddiqui MM, Schuch G, Pohl HG, Fuhr J, Perin L, Soker S, Atala A (2005) Propagation, expansion, and multilineage differentiation of human somatic stem cells from dermal progenitors. Stem Cells Dev 14(3):337–348.  https://doi.org/10.1089/scd.2005.14.337CrossRefPubMedGoogle Scholar
  13. 13.
    Chen FG, Zhang WJ, Bi D, Liu W, Wei X, Chen FF, Zhu L, Cui L, Cao Y (2007) Clonal analysis of nestin(−) vimentin(+) multipotent fibroblasts isolated from human dermis. J Cell Sci 120(Pt 16):2875–2883.  https://doi.org/10.1242/jcs.03478CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Hua Qian
    • 1
  • Xue Leng
    • 1
  • Jie Wen
    • 1
  • Qian Zhou
    • 1
  • Xin Xu
    • 1
  • Xunwei Wu
    • 1
    Email author
  1. 1.Laboratory for Tissue Engineering and Regeneration, and Shandong Provincial Key Laboratory of Oral Tissue Regeneration, School of StomatologyShandong UniversityJinanChina

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