Epidermal cells adhere preferentially to type IV (basement membrane) collagen.

Abstract

Epidermal cells from adult guinea pig skin attach and differentiate preferentially on substrates of type IV (basement membrane) collagen, compared to those of types I-III collagen. In contrast, guinea pig dermal fibroblasts attach equally well to all four collagen substrates. Fibronectin mediates the attachment of fibroblasts but not of epidermal cells to collagen. KEY WORDS epidermal cells adhesion collagen -basement membrane differentiationSeveral chemically and genetically distinct collagens are found in mammalian species. Types I and III collagen are widely distributed (14). Type II collagen is found only in cartilage (13) and type IV in basement membranes (9). The matrices that these collagens form differ in appearance and physical characteristics. Collagen substrates enhance the attachment (10), growth (2), and differentiation (5) of various cell types. Such observations suggest that the interaction of cells with collagen may have important functions in vivo. Some clue to the mechanism(s) operative in this interaction came from recent studies (10, 11) which indicate that fibroblasts do not bind directly to collagen substrates but that a membrane protein (the collagen-cen attachment protein) mediates the attachment of cells to collagen substrates. This protein (or proteins) is similar or identical to CSP (23), LETS protein (7) and cold insoluble globulin (15,19), and has been named fibronectin (18). Fetal calf serum used in cell culture is a rich source of fibronectin (10).It seemed conceivable to us that collagen-cell interactions depend not only on mediator proteins but also on either the chemical characteristics of the collagen substrate or on the nature of the attaching cell. To test this hypothesis, we compared the attachment of two functionally diverse cell types, fibroblasts and epidermal cells, to various collagen substrates and their dependence on fibronectin for the mediation of attachment. In contrast to fibroblasts, epidermal cells from postembryonic animals are difficult to culture in vitro unless conditioned media and collagen substrates (8) or feeder layers of fibroblasts (17) are used. We show here that epidermal cells differ from fibroblasts in their attachment to collagen substrates and that epidermal cells attach best to type IV collagen whereas fibroblasts attach equally well to all collagen types. Further, we demonstrate that fibronectin enhances the attachment of fibroblasts to collagen but does not affect epidermal cell attachment. MATERIALS AND METHODS Collagen SubstratesType I collagen was prepared from the skins of lathyritic rats (1), type II from a rat chondrosarcoma (20), type III from fetal calf skin (4), and type IV from a murine sarcoma (16,22). The purity and identity to standards of these collagens was confn'med by sodium dodecyl sulfate (SDS) gel electrophoresis (12) and amino acid analysis. The various collagens (1 mg/ml in 0.5 M acetic acid) were stored at -20"C. To study the attachment of cells, the stock solutions of collagen were diluted to 10 /~g/ml with wate...

The different effects of type I and IV collagens on the survival and proliferation of cultured BSC-1 cells.

“…A similar effect on spreading has been reported in the case of various lines of cultured cells, in particular, epithelial cells (7,10,15). The augmented spreading was observed with the fraction of more than 100 kDa, but not with the fraction of less than 100 kDa.…”

“…Type I collagen is the natural substratum for fibroblasts (11), as is type IV collagen for epithelial cells (15). Type I collagen, but not type IV collagen, promotes morphological changes in and dissociation of epithelial cells (19).…”

Mammalian collagen IV

“…This interaction is critical for a variety of biological processes, including cell adhesion, migration, survival, proliferation, and differentiation. Cell culture studies have shown that collagen IV is the binding substrate for a large number of cell types including platelets (Santoro, 1986;Staatz et al, 1990), hepatocytes (Rubin et al, 1981), keratinocytes (Murray et al, 1979), endothelial (Cheng and Kramer, 1989;Herbst et al, 1988), mesangial (Setty et al, 1998), pancreatic (Kaido et al, 2004), and tumor cells such as breast and prostate carcinoma (Abecassis et al, 1987;Dedhar et al, 1993), melanoma (Chelberg et al, 1989), fibrosarcoma, and glioma (Aumailley and Timpl, 1986;Knight et al, 2000). Cell attachment to collagen IV is mediated by multiple binding sites within both triple-helical and NC1 domains, suggesting involvement of several adhesion receptors (Chelberg et al, 1989;Herbst et al, 1988;Wayner and Carter, 1987).…”

Culture of prostate epithelial cells of the rhesus monkey on extracellular matrix substrate: influence of steroids and insulin-like growth factors

“…The major components of ECM are collagen, elastin, proteoglycans, glycoprotein fibrils and anchorage proteins such as fibronectin and laminin and its composition varies in different tissues (Kleinman et al 1981). Murray et al (1979) also reported a four-to fivefold increase in the attachment of epidermal cells in collagen IV-coated flasks than in flasks coated with types I-III collagen. Collagen substrate enhances the attachment, growth and differentiation of various cell types (Ehrman & Gey 1956, Klebe 1974 and cells attach to collagen or laminin substrates not directly but through membrane proteins like fibronectin (Rouslahti et al 1973).…”