Summary
The effects of collagenous substrata, fibronectin, and fetal bovine serum on the adhesion, proliferation, and adipogenesis of rat stromal-vascular cells are reported. There was no effect on initial stromal-vascular cell-attachment by fetal bovine serum or fibronectin. The number of cells attached to a hydrated collagen-gel was almost twice (P<0.04) the number attached to dried collagen-gel or dried denatured collagen-gel. Total number of cells after 5 days in culture was similar among the collagenous substrata and among the treatments with or without fibronectin in the growth media. Total number of cells increased significantly (P<0.02) with 10% FBS. Adipocytic formation was inhibited by hydrated collagen-gel (P<0.02) compared to dried collagen-gel or dried, denatured collagenous substrata. An interaction occurred between dried, denatured gel and fetal bovine serum so that total formation of adipocytes increased by increasing the level of fetal bovine serum (P<0.07). Adipocytic formation was inhibited by hydrated collagen-gel at all levels of fetal bovine serum. The percentage of cells that converted to adipocytes was significantly lower (P<0.01) on hydrated collagen-gel compared to dried, denatured or dried collagen-gel. Percentage of conversion was not significantly different among levels of fetal bovine serum, although this percentage increased as fetal bovine serum level increased. Adipocytic conversion was not different between fibronectin-treated or untreated cells. Morphology of stromal vascular cells was similar on dried collagen and dried, denatured collagen-gel, but tended to remain bipolar on hydrated collagen-gel. These studies indicate that fetal bovine serum in combination with the extracellular matrix (dried, denatured collagen) increased the differentiation of rat stromal-vascular cells into adipocytes, and that hydrated collagen inhibited differentiation.
Similar content being viewed by others
References
Bjorntorp P, Karlsson M, Petterson P, Sypniewska G (1980) Differentiation and function of rat adipocyte precursor cells in primary culture. J Lipid Res 21:714–723
Delvos V, Gajdisek C, Sage H, Harker LA, Scwartz SM (1982) Interactions of vascular wall cells with collagen gels. Lab Invest 48:61–72
Elsdale T, Bard J (1972) Collagen substrata for studies on cell behavior. J Cell Biol 54:626–637
Giancotti F, Tarone G, Knudsen K, Damsky C, Comogleo PM (1985) Cleavage of a 135 Kd cell surface glycoprotein correlates with loss of fibroblast adhesion to fibronectin. Exp Cell Res 156:182–190
Greenburg G, Hay ED (1986) Cytodifferentiation and tissue phenotype change during transformation of embryonic lens epithelium to mesenchyme-like cells in vitro. Dev Biol 115:363–379
Grinnell FG, Bennett MH (1981) Fibroblast adhesion on collagen substrata in the presence and absence of plasma fibronectin. J Cell Sci 48:19–34
Hausman GJ (1981) Techniques for studying adipocytes. Stain Technol 56:149–154
Hausman GJ, Novakofski J, Martin RJ, Thomas GB (1984) The histochemistry of developing adipocytes in primary stromalvascular cultures of rat adipose tissue. Histochemistry 80:353–358
Humason GL (1982) Animal tissue techniques. Freeman, San Francisco, p 15
Klebe RJ (1974) Isolation of a collagen-dependent cell attachment factor. Nature 250:248–251
Kleinman HK, Klebe RJ, Martin GR (1981) Role of collagen matrices in the adhesion and growth of cells. J Biol Chem 88:473–485
Kleinman HK, McGoodwin EB, Martin GR, Klebe RJ, Fietzek PP, Wooley DE (1978) Localization of the binding site for cell attachment protein in the α 1(1) chain of collagen. J Biol Chem 253:5642–5646
Mosher DF (1980) Fibronectin. Prog Hemost Thromb 5:111–151
Pytela R, Pierschbacher MD, Rusolahti E (1985) Identification of a 140 Kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell 40:191–198
Richardson RL, Hausman GJ, Campion DR, Thomas GB (1986) Adipocyte development in primary rat cell cultures: A scanning electron microscopy study. Anat Rec 216:144–150
Rubin K, Oldberg A, Hook M, Obrink B (1978) Adhesion of rat hepatocytes to collagen. Exp Cell Res 117:165–177
SAS (1985) SAS user's guide, Statistical Analysis Systems, Inc., Cary, NC
Schor SL, Court J (1979) Different mechanisms in the attachment of cells to native and denatured collagen. J Cell Sci 38:267–281
Smith JC, Singh JP, Lillquist JS, Goon DS, Stiles CD (1982) Growth factors adherent to cell substrata are mitogenically active in situ. Nature 296:154–156
Spiegelman BM, Ginty CA (1983) Fibronectin modulation of cell shape and lipogenic gene expression in 3T3-adipocytes. Cell 35:657–666
Terranova VP, Aumailley M, Sulton LH, Martin GR, Kleinman HK (1986) Regulation of cell attachment and cell number by fibronectin and laminin. J Cell Physiol 127:473–479
Yamada KM (1982) Fibronectin and other structural proteins. In: Hay ED (ed) Cell biology of the extracellular matrix. Plenum Press, New York, pp 95–114
Yoshizato K, Taira T, Yamamoto N (1985) Growth inhibition of human fibroblasts by reconstituted collagen fibrils. Biomed Res 6:61–72
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Richardson, R.L., Campion, D.R. & Hausman, G.J. Adhesion, proliferation, and adipogenesis in primary rat cell cultures: effects of collagenous substrata, fibronectin, and serum. Cell Tissue Res. 251, 123–128 (1988). https://doi.org/10.1007/BF00215456
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00215456