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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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, 39,027ā39,031.
Heughan, C. and Hunt, T. (1975) Some aspects of wound healing research: a review. Can. J. Surg. 18, 118ā126.
Gabbiani, G. (2003) The myofibroblast in wound healing and fibrocontractive diseases. J. Pathol. 200, 500ā503.
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, 880ā889.
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, 397ā414.
RittiĆ©, L. and Fisher, G. (2002) UV-light-induced signal cascades and skin aging. Ageing Res. Rev. 1, 705ā720.
Hiebert, C. (1959) The carcinogenic action of methylcholanthrene on mouse skin in organ culture. Cancer 12, 663ā672.
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, 505ā510.
Hayflick, L. and Moorhead, P. (1961) The serial cultivation of human diploid cell strains. Exp. Cell Res. 25, 585ā621.
Cristofalo, V. and Pignolo, R. (1993) Replicative senescence of human fibroblastlike cells in culture. Physiol. Rev. 73, 617ā638.
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, 1274ā1278.
Grinnell, F. (2003) Fibroblast biology in three-dimensional collagen matrices. Trends Cell Biol. 13, 264ā269.
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, 129ā133.
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, 493ā503.
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, 301ā305.
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, 365ā374.
Arora, P., Narani, N., and McCulloch, C. (1999) The compliance of collagen gels regulates transforming growth factor-Ć induction of a-smooth muscle actin in fibroblasts. Am. J. Pathol. 154, 871ā882.
Kessler, D., Dethlefsen, S., Haase, I., et al. (2001) Fibroblasts in mechanically stressed collagen lattices assume a āsyntheticā phenotype. J. Biol. Chem. 276, 36,575ā36,585.
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, 283ā291.
Freeman, A., Igel, H., Herrman, B., and Kleinfeld, K. (1976) Growth and characterization of human skin epithelial cell cultures. In Vitro 12, 352ā362.
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, 1052ā1054.
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, 341ā352.
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, 161ā174.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights 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