Growth of human foreskin fibroblasts in a serum‐free, defined medium without platelet‐derived growth factor

Abstract

The hormones which support growth, in vitro, of normal, neonatal human foreskin fibroblasts were determined. Whereas thrombin and hydrocortisone were major growth stimulants, platelet-derived growth factor was not. Human foreskin fibroblasts grew in a serum-free, biochemically defined medium consisting of epidermal growth factor (100 ng/ml), insulin (100 ng/ml), transferrin 10 micrograms/ml), thrombin (1 microgram/ml), ascorbic acid (10 micrograms/ml), and hydrocortisone (5 x 10(-5) M) in a 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F-12, supplemented with ovalbumin (1 mg/ml) and trace elements. The growth achieved was comparable to that achieved with 5% fetal bovine serum. Neither platelet-derived growth factor, fibroblast growth factor, nor somatomedin activity increased proliferation. This serum-free medium, designated Defined Medium F, provides a biochemically defined system for growth and limited subcultivation of human foreskin fibroblasts in vitro.

Interaction of native and cell-modified low density lipoprotein with collagen gel.

“…Porcine aortic SMC were grown from explants of the middle layers of media from the arch region. 47 Cultures of human skin fibroblasts (HSF) were established from single cell suspensions obtained by enzymatic dispersion of normal foreskin dermis 48 of patients ages 7 months to 30 years. Stock cultures of all cell types were grown in 100 mm tissue culture dishes in Dulbecco's modified Eagle's medium (DMEM) (Flow Laboratories, Incorporated, Mississauga, Ontario) supplemented with 1 mM nonessential amino acids (GIBCO Canada, Burlington, Ontario) and sodium bicarbonate (2.25 g/1).…”

“…SMC received 10% (vol/vol) fetal bovine serum (FBS) instead of the special calf serum. In addition, after the first day in primary culture, EC received EC growth factor in the form of a crude extract of bovine brain 48 at 150 (jug protein/ml medium. For EC and SMC, culture dishes were coated with human plasma fibronectin as previously described.…”

[125I]EGF binding ability is stable throughout the replicative life‐span of WI‐38 cells

“…We have recently reported Cristofalo, 1980, 1981) on the development of such a serumfree medium for the WI-38 cell line (normal human embryonic lung-derived fibroblast-like cells). A similar medium has been independently developed for an analogous cell line (Walthall and Ham, 1981) and more recently, a slightly modified medium for human foreskin fibroblasts (Weinstein et al, 1982). Our serum-free medium consists of basal medium MCDB-104 supplemented with a partially purified preparation of plateletderived growth factor (PDGF), epidermal growth factor (EGF), insulin (INS), transferrin (TRS), and dexamethasone (DEX).…”

Growth Factors and Wound Healing in the Hamster

“…We then added to wounds other preparations known to enhance the growth of fibroblasts. Purified human α‐thrombin (10 μg per application), defined medium F for fibroblasts 14 (20 μ1 per application), and liver cell supernatant 16 (30.8 μg per application) also failed to accelerate wound healing (Table 1).…”

“…Defined medium F was a modification of that of Weinstein and colleagues 14 and contained EGF (100 ng/ml), INS (100 ng/ml), transferrin (10 μg/ml), thrombin (1 μg/ml), DEX (50 ng/ml), ovalbumin (1 mg/ml), ascorbic acid (10 μg/ml), and trace elements 15 in a 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F‐12 medium.…”