title: Culture medium and cell density impact gene expression in normal skin and abnormal scar-derived fibroblasts.
abstract: Fibroblasts, the main cell type of the dermis, are responsible for production and remodeling of extracellular matrix during wound healing. Disruption of either production or degradation of extracellular matrix can lead to abnormal scarring, resulting in hypertrophic scar or keloid scar. Aberrations in proliferation and gene expression have been observed in fibroblasts isolated from abnormal scars, but differences observed may be related to biologic responses to growth conditions and media formulations. This study examined gene expression in primary human fibroblasts from normal skin or abnormal scar in two culture media formulations and three relative cell densities. In general, higher expression of collagen type 1 alpha-1 (COL1A1) and alpha-2 (COL1A2) and matrix metalloproteinase 3 (MMP3) and lower levels of MMP1 were observed in all cell strains cultured in standard medium containing 10% fetal bovine serum compared with cells cultured in medium optimized for proliferation. Normal and scar-derived fibroblasts exhibited differences in gene expression in specific response to media formulations and cell density. COL1A1 and COL1A2 were increased, and MMP1 and MMP3 were decreased, in keloid cells compared with normal fibroblasts under most conditions analyzed. However, expression of plasminogen activator inhibitor 1 in keloid fibroblasts, which was significantly different than in normal fibroblasts, was either increased or decreased in response to the medium formulation and relative cell density. A related gene, plasminogen activator inhibitor 2, was shown for the first time to be significantly increased in keloid fibroblasts compared with normal fibroblasts, in both media formulations and at all three cell densities. The results emphasize the critical role of culture conditions in interpretation of cell behavior and expression data and for comparison of cells representing normal and fibrotic phenotypes.
authors: Research Department, Shriners Hospitals for Children, Cincinnati, Ohio 45229, USA.
authors: Kevin L
authors: KL
authors: McFarland
authors: Kathryn
authors: K
authors: Glaser
authors: Jennifer M
authors: JM
authors: Hahn
authors: Steven T
authors: ST
authors: Boyce
authors: Dorothy M
authors: DM
authors: Supp
mesh_terms: D000818:Animals; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D017439:Cicatrix, Hypertrophic / Q000378:metabolism* / Q000473:pathology; D024042:Collagen Type I / Q000378:metabolism*; D000090602:Collagen Type I, alpha 1 Chain; D003470:Culture Media / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D044967:Serum / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D003470:Culture Media; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1