Skip to main content

Isolation and Culture of Skin Fibroblasts

  • Protocol
Fibrosis Research

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 117))

Abstract

Dermal fibroblasts synthesize and organize skin connective tissue (dermis). Fibroblast cultures provide a powerful tool for investigating both the molecular mechanisms that regulate skin connective tissue production and the pathophysiology of fibrotic skin diseases. In this chapter, we describe detailed procedures for establishing and maintaining primary cultures of adult human fibroblasts. We also provide protocols for constructing three-dimensional dermal and skin equivalent culture models.

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. Verzijl, N., DeGroot, J., Thorpe, S., et al. (2000) Effect of collagen turnover on the accumulation of advanced glycation end products. J. Biol. Chem. 275, 39,027ā€“39,031.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  2. Heughan, C. and Hunt, T. (1975) Some aspects of wound healing research: a review. Can. J. Surg. 18, 118ā€“126.

    PubMedĀ  CASĀ  Google ScholarĀ 

  3. Gabbiani, G. (2003) The myofibroblast in wound healing and fibrocontractive diseases. J. Pathol. 200, 500ā€“503.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  4. LeRoy, E. (1974) Increased collagen synthesis by scleroderma skin fibroblasts in vitro: a possible defect in the regulation or activation of the scleroderma fibroblast. J. Clin. Invest. 54, 880ā€“889.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  5. Jimenez, S. and Saitta, B. (2000) Alternations in the regulation of expression of the Ī±1(I) collagen gene (COL1A1) in systemic sclerosis (scleroderma). Springer Semin. Immunopathol. 21, 397ā€“414.

    Google ScholarĀ 

  6. RittiĆ©, L. and Fisher, G. (2002) UV-light-induced signal cascades and skin aging. Ageing Res. Rev. 1, 705ā€“720.

    ArticleĀ  PubMedĀ  Google ScholarĀ 

  7. Hiebert, C. (1959) The carcinogenic action of methylcholanthrene on mouse skin in organ culture. Cancer 12, 663ā€“672.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  8. Aumailley, M., Krieg, T., Razaka, G., MĆ¼ller, P., and Bricaud, H. (1982) Influence of cell density on collagen biosynthesis in fibroblast cultures. Biochem. J. 206, 505ā€“510.

    PubMedĀ  CASĀ  Google ScholarĀ 

  9. Hayflick, L. and Moorhead, P. (1961) The serial cultivation of human diploid cell strains. Exp. Cell Res. 25, 585ā€“621.

    ArticleĀ  Google ScholarĀ 

  10. Cristofalo, V. and Pignolo, R. (1993) Replicative senescence of human fibroblastlike cells in culture. Physiol. Rev. 73, 617ā€“638.

    PubMedĀ  CASĀ  Google ScholarĀ 

  11. Bell, E., Ivarsson, B., and Merrill, C. (1979) Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc. Natl. Acad. Sci. USA 76, 1274ā€“1278.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  12. Grinnell, F. (2003) Fibroblast biology in three-dimensional collagen matrices. Trends Cell Biol. 13, 264ā€“269.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  13. Eckes, B., Mauch, C., HĆ¼ppe, G., and Krieg, T. (1993) Downregulation of collagen synthesis in fibroblasts within three-dimensional collagen lattices involves transcriptional and posttranscriptional mechanisms. FEBS 318, 129ā€“133.

    ArticleĀ  CASĀ  Google ScholarĀ 

  14. Mauch, C., Hatamochi, A., Scharffetter, K., and Krieg, T. (1988) Regulation of collagen synthesis in fibroblasts within a three-dimensional collagen gel. Exp. Cell Res. 178, 493ā€“503.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  15. Mauch, C., Adelmann-Grill, B., Hatamochi, A., and Krieg, T. (1989) Collagenase gene expression in fibroblasts is regulated by a three-dimensional contact with collagen. FEBS 250, 301ā€“305.

    ArticleĀ  CASĀ  Google ScholarĀ 

  16. Seltzer, J., Lee, A.-Y., Akers, K., et al. (1994) Activation of 72-kDa type IV collagenase/gelatinase by normal fibroblasts in collagen lattices is mediated by integrin receptors but is not related to lattice contraction. Exp. Cell Res. 213, 365ā€“374.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  17. Arora, P., Narani, N., and McCulloch, C. (1999) The compliance of collagen gels regulates transforming growth factor-Ɵ induction of a-smooth muscle actin in fibroblasts. Am. J. Pathol. 154, 871ā€“882.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  18. Kessler, D., Dethlefsen, S., Haase, I., et al. (2001) Fibroblasts in mechanically stressed collagen lattices assume a ā€œsyntheticā€ phenotype. J. Biol. Chem. 276, 36,575ā€“36,585.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  19. Roguet, R., Cohen, C., Dossou, K., and Rougier, A. (1994) Episkin, a constituted human epidermis for assessing in vitro the irritancy of topically applied compounds. Toxicol. Vitro 8, 283ā€“291.

    ArticleĀ  CASĀ  Google ScholarĀ 

  20. Freeman, A., Igel, H., Herrman, B., and Kleinfeld, K. (1976) Growth and characterization of human skin epithelial cell cultures. In Vitro 12, 352ā€“362.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  21. Bell, E., Ehrlich, H., Buttle, D., and Nakatsuji, T. (1981) Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness. Science 211, 1052ā€“1054.

    ArticleĀ  PubMedĀ  CASĀ  Google ScholarĀ 

  22. Pouliot, R., Germain, L., Auger, F., Tremblay, N., and Juhasz, J. (1999) Physical characterization of the stratum corneum of an in vitro human skin equivalent produced by tissue engineering and its comparision with normal human skin by ATRFTIR spectroscopy and thermal analysis (DSC). Biochim. Biophys. Acta 1439, 341ā€“352.

    PubMedĀ  CASĀ  Google ScholarĀ 

  23. Nickoloff, B., Mitra, R., Fisher, G., and Voorhees, J. (1989) Decreased growth inhibition by recombinant gamma interferon is associated with increased transforming growth factor-Ī± production in keratinocytes cultured from psoriatic lesions. Brit. J. Dermatol. 121, 161ā€“174.

    ArticleĀ  CASĀ  Google ScholarĀ 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

Ā© 2005 Humana Press Inc.

About this protocol

Cite this protocol

RittiƩ, L., Fisher, G.J. (2005). Isolation and Culture of Skin Fibroblasts. In: Varga, J., Brenner, D.A., Phan, S.H. (eds) Fibrosis Research. Methods in Molecular Medicine, vol 117. Humana Press. https://doi.org/10.1385/1-59259-940-0:083

Download citation

  • DOI: https://doi.org/10.1385/1-59259-940-0:083

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-479-1

  • Online ISBN: 978-1-59259-940-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics