Regulation of collagen metabolism and cell growth by epidermal growth factor and ascorbate in cultured human skin fibroblasts


Correspondence to R. Hata, Department of Tissue Physiology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan 101


Epidermal growth factor (2–50 ng/ml), prepared from mouse submaxillary glands, stimulated growth and the synthesis of non-collagenous proteins and hyaluronic acid, but inhibited collagen synthesis in cultured human skin fibroblasts, both stimulation and inhibition being dose-dependent. All these effects may be intrinsic functions of the epidermal growth factor molecule, because these effects were cancelled by the co-presence of antiserum specific for epidermal growth factor and because they were also observed following the addition of human epidermal growth factor produced urogastrone cDNA. On the other hand, l-ascorbate (vitamin C) stimulated growth and collagen synthesis, as well as synthesis of non-collagenous proteins, with no significant effect on hyaluronic acid synthesis.

Co-presence of epidermal growth factor and ascorbate gave additive effects on growth and protein synthesis of the cells. These results suggest that the two growth-promoting factors, epidermal growth factor and l-ascorbate, modulate metabolism of extracellular matrix components as well as cell growth in a quite different manner in human skin fibroblasts.


dibutyryl-adenosine 3′,5′-monophosphate


Dulbecco's modified Eagle's medium supplemented with Fungizone (250 μg/l), penicillin G (50 mg/l), dihydrostreptomycin (50 mg/l) and Hepes (3 g/l)


DMEM containing 10% fetal bovine serum


epidermal growth factor


human epidermal growth factor


mouse epidermal growth factor


0.05 M Tris/HCl containing 0.15 M NaCl, pH 7.4


Clostridium histolyticum collagenase (EC


lysyl hydroxylase (EC


pepsin (EC


prolyl hydroxylase (EC


pronase (EC