Serum glucocorticoids have persistent and controlling effects on insulinlike growth factor i action under serum-free assay conditions in cultured human fibroblasts

Summary The biological actions of insulinlike growth factor I (IGF-I) measured under serum-free assay conditions were found to be significantly influenced by prior subculture conditions for adult human fibroblasts. Glucocorticoids seemed to be the major medium variable affecting IGF-I action. IGF-I added to serum-free cultures had little or no effect on ¹⁴C]aminoisobutyric acid uptake or ³H]thymidine incorporation in human fibroblasts previously maintained in media containing serum with low glucocorticoid levels or in serum stripped of endogenous steroids. However, a 48-h preincubation with dexamethasone resulted in a marked synergistic increase in IGF-I stimulation of ¹⁴C]aminoisobutyric acid uptake and ³Hthymidine incorporation in these cultures. In contrast, IGF-I in serum-free medium seemed to be a potent mitogenic and metabolic stimulus for human fibroblasts which had been subcultured in media with a high glucocorticoid content, either endogenous, or supplemented. After these culture conditions, a 48-h preexposure to dexamethasone had no further enhancing effect on IGF-I action. Dexamethasone also potentiated IGF-I, insulin, and epidermal growth factor stimulation of fibroblast replication depending on the earlier subcultivation conditions. Thus, glucocorticoids are important modulators of IGF-I bioactivity in cultured human fibroblasts. Serum glucocorticoids can exert a profound influence on the biological phenomena measured in cell culture, even when the serum has been removed before the actual experiment, and must be carefully taken into account for accurate evaluation of the biological function of IGF-I and other growth factors. This work was supported in part by grants DK28229, DK36054, CA42106, RCDA01275, and DK24085 from the National Institutes of Health, Bethesda, MD.