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.
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 188.8.131.52)
lysyl hydroxylase (EC 184.108.40.206)
pepsin (EC 220.127.116.11)
prolyl hydroxylase (EC 18.104.22.168)
pronase (EC 22.214.171.124)