Skip to main content

Isolation and Cultivation of Human Scalp Interfollicular Epidermal Stem Cells

  • Protocol
  • First Online:
Skin Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 989))

  • 2957 Accesses

Abstract

Skin regeneration is intricately controlled by epidermal stem cells. In human skin, the long-lived, slow-cycling, and highly proliferative stem cells are located in the basal layer of the interfollicular epidermis (IFE). The ability to isolate and culture human IFE stem cells (IFESCs) offers fascinating therapeutic potential for skin diseases as well as epithelial tissue engineering. Here we describe a straightforward strategy for generation of β1 integrin+/CD24 IFESCs from scalp with defined, serum-free, feeder-free medium and collagen I-coated culture plates. The use of defined media throughout isolation and cultivation allows for detailed investigation of the molecular events involved in ESC self-renewal and differentiation as well as provides a safe source for clinical use.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Quesenberry P, Levitt L (1979) Hematopoietic stem cells (second of three parts). N Engl J Med 301:819–823

    Article  PubMed  CAS  Google Scholar 

  2. Lajtha LG (1979) Stem cell concepts. Differentiation 14:23–34

    Article  PubMed  CAS  Google Scholar 

  3. Ghazizadeh S, Taichman LB (2005) Organization of stem cells and their progeny in human epidermis. J Invest Dermatol 124:367–372

    Article  PubMed  CAS  Google Scholar 

  4. Lavker RM, Sun TT (1982) Heterogeneity in epidermal basal keratinocytes: morphological and functional correlations. Science 215:1239–1241

    Article  PubMed  CAS  Google Scholar 

  5. Lavker RM, Sun TT (2000) Epidermal stem cells: properties, markers, and location. Proc Natl Acad Sci USA 97:13473–13475

    Article  PubMed  CAS  Google Scholar 

  6. Levy V, Lindon C, Zheng Y et al (2007) Epidermal stem cells arise from the hair follicle after wounding. FASEB J 21:1358–1366

    Article  PubMed  CAS  Google Scholar 

  7. Ghazizadeh S, Taichman LB (2001) Multiple classes of stem cells in cutaneous epithelium: a lineage analysis of adult mouse skin. EMBO J 20:1215–1222

    Article  PubMed  CAS  Google Scholar 

  8. Ito M, Liu Y, Yang Z et al (2005) Stem cells in the hair follicle bulge contribute to wound repair but not to homeostasis of the epidermis. Nat Med 11:1351–1354

    Article  PubMed  CAS  Google Scholar 

  9. Levy V, Lindon C, Harfe BD et al (2005) Distinct stem cell populations regenerate the follicle and interfollicular epidermis. Dev Cell 9:855–861

    Article  PubMed  CAS  Google Scholar 

  10. Owens DM, Watt FM (2003) Contribution of stem cells and differentiated cells to epidermal tumours. Nat Rev Cancer 3:444–451

    Article  PubMed  CAS  Google Scholar 

  11. Rheinwald JG, Green H (1975) Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6:331–343

    Article  PubMed  CAS  Google Scholar 

  12. 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 3:713–724

    Article  Google Scholar 

  13. Roh C, Roche M, Guo Z et al (2008) Multi-potentiality of a new immortalized epithelial stem cell line derived from human hair follicles. In Vitro Cell Dev Biol Anim 44:236–244

    Article  PubMed  CAS  Google Scholar 

  14. Jones PH, Harper S, Watt FM (1995) Stem cell patterning and fate in human epidermis. Cell 80:83–93

    Article  PubMed  CAS  Google Scholar 

  15. Ji L, Allen-Hoffmann BL, de Pablo JJ et al (2006) Generation and differentiation of human embryonic stem cell-derived keratinocyte precursors. Tissue Eng 12:665–679

    Article  PubMed  CAS  Google Scholar 

  16. Lorenz K, Rupf T, Salvetter J et al (2009) Enrichment of human beta 1 bri/alpha 6 bri/CD71 dim keratinocytes after culture in defined media. Cells Tissues Organs 189:382–390

    Article  PubMed  CAS  Google Scholar 

  17. Jiang S, Zhao L, Purandare B et al (2010) Differential expression of stem cell markers in human follicular bulge and interfollicular epidermal compartments. Histochem Cell Biol 133:455–465

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The protocols described here were developed under the support of a generous gift from the Minnesota Jewish Foundation (2006) and an ASDS Cutting Edge Grant (BMH). The authors thank Shu Jiang, M.D., Ph.D. for her technical assistance.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media New York

About this protocol

Cite this protocol

Zhao, L., Hantash, B.M. (2013). Isolation and Cultivation of Human Scalp Interfollicular Epidermal Stem Cells. 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_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-330-5_2

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-329-9

  • Online ISBN: 978-1-62703-330-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics