Keratin 19 as a Stem Cell Marker In Vivo and In Vitro

  • Danielle Larouche
  • Cindy Hayward
  • Kristine Cuffley
  • Lucie Germain
Part of the Methods in Molecular Biology™ book series (MIMB, volume 289)


The skin is a dynamic tissue in which terminally differentiated keratinocytes are replaced by the proliferation of new epithelial cells that will undergo differentiation. The rapid and continual turnover of skin throughout life depends on a cell population with unique characteristics: the stem cells. These cells are relatively undifferentiated, retain a high capacity for self-renewal throughout their lifetime, have a large proliferative potential, and are normally slow cycling. The long-term regeneration of grafted cultured epidermis indicates that epidermal stem cells are maintained in cultures. In animals they can be identified with 3H-thymidine or bromodeoxyuridine based on their property of slow cycling. The development of markers such as keratin 19 also permits their study in human tissues. In this chapter, protocols to study skin stem cells using their property of slow cycling and their expression of keratin 19 will be described in detail. The methods include the double labeling of tissues for keratin 19 and label-retaining cells (autoradiography of 3H-thymidine) in situ. The labeling of keratin 19 by immunofluorescence of by flow cytometry is described for cells in vitro.

key Words

Stem cells keratin keratin 19 epidermis hair follicles human skin flow cytometry 


  1. 1.
    Green, H., Kehinde, O., and Thomas, J. (1979) Growth of cultured human epidermal cells into a multiple epithelia suitable for grafting. Proc. Natl. Acad. Sci. USA 76, 5665–5668.PubMedCrossRefGoogle Scholar
  2. 2.
    Germain, L., Rouabhia, M., Guignard, R., Carrier, L., Bouvard, V., and Auger, F A. (1993) Improvement of human keratinocyte isolation and culture using thermolysin. Burns 2, 99–104.CrossRefGoogle Scholar
  3. 3.
    Germain, L., Michel, M., Fradette, J., Xu, W., Godbout, M-J., and Li, H. (1997) Skin stem cell identification and culture: a potential tool for rapid epidermal sheet production and grafting, in Skin Substitute Production by Tissue Engineering: Clinical and Fundamental Applications. (Rouabhia, M., ed.), Landes Bioscience, Austin, TX, pp. 177–210.Google Scholar
  4. 4.
    Michel, M., Török, N., Godbout, M.-J., Lussier, M., Gaudreau, P., Royal, A. et al. (1996) Keratin 19 as a biochemical marker of skin stem cells in vivo and in vitro: keratin 19 expressing cells are differentially localized in function of anatomic sites, and their number varies with donor age and culture stage. J. Cell. Sci. 109, 1017–1028.PubMedGoogle Scholar
  5. 5.
    Michel, M., ĽHeureux, N., Auger, F. A., and Germain, L. (1997) From newborn to adult: phenotypic and functional properties of skin equivalent and human skin as a function of donor age. J. Cell Phys. 171, 179–189.CrossRefGoogle Scholar
  6. 6.
    Fradette, J., Larouche, D., Fugère, C., Guignard, R., Beauparlant, A., Couture, V., et al. (2003) Normal human Merkel cells are present in epidermal cell populations isolated and cultured from glabrous and hairy skin sites. J. Invest. Dermatol. 120, 313–317.PubMedCrossRefGoogle Scholar
  7. 7.
    Lussier, M., Ouellet, T., Lampron, C., Lapointe, L., and Royal, A. (1989) Mouse keratin 19: complete amino acid sequence and gene expression during development. Gene 85, 435–444.PubMedCrossRefGoogle Scholar
  8. 8.
    Cotsarelis, G., Sun, T. T., and Lavker, R. M. (1990) Label-retaining cells reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis. Cell 61, 1329–1337.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • Danielle Larouche
    • 1
  • Cindy Hayward
    • 1
  • Kristine Cuffley
    • 1
  • Lucie Germain
    • 1
  1. 1.Laboratoire de Recherche des Grands Brüles/LOEX and Department of SurgeryLaval UniversityQuébecCanada

Personalised recommendations