Glucocorticoid receptors and inhibition of neonatal mouse dermal fibroblast growth in primary culture

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Abstract
Summary Primary cultures of dermal fibroblasts from neonatal mice were used to investigate some of the anti-inflammatory effects of glucocorticoids in vitro as influenced by the genetic background of two different strains of mice (A/J and C57 B1/6J). Fibroblasts were cultured in the absence or presence of various glucocorticoids for 2–7 days. After 4–7 days in the presence of steroid, DNA synthesis was reduced by 50–85% while protein synthesis was inhibited by 50–60%. Corticosterone produced a dose-dependent inhibition of DNA synthesis in these cells with a 50% reduction occurring at 10 nM. Specific, high affinity, low capacity binding proteins for [3H]dexamethasone or [3H]triamcinolone acetonide were identified in the cytoplasm of neonatal dermal fibroblasts which had an apparent Kd of 9 nM and ∼5,200–6,400 binding sites/cell. Sedimentation analysis of the [3H]triamcinolone acetonide-receptor complexes on low salt glycerol gradients exhibited binding in the 7 to 8 S region of the gradients. These studies demonstrate that inhibition of growth of primary cultures of mouse neonatal dermal fibroblasts by glucocorticoids is probably mediated by a receptor-mediated pathway, and that this primary culture system might be useful in delineating other anti-inflammatory effects of glucocorticoids in vitro.