Regulation of insulin binding and stimulation of sugar transport in cultured human fibroblasts by sugar levels in the culture medium.

Studies were carried out on cultures of human skin fibroblasts to explore the effects of culture medium glucose levels on insulin binding and action. Cell cultures in 5.55 mM glucose-containing medium depleted their medium glucose within 3 days, and at that time exhibited elevated deoxy-D-glucose (2-DG) transport (84% greater than control cultures fed 22.2 mM glucose) and failure of insulin to stimulate 2-DG transport (an insulin:control transport ratio of 1.02). There was also a significant negative correlation between basal 2-DG transport and insulin binding (r = -0.621; n = 29; P less than 0.01), while insulin binding exhibited a significant positive correlation with insulin action (r = 0.816; n = 12; P less than 0.01). Glucose starvation of cultures for 18 h resulted in several changes: a 49% decrease in specific 125I-insulin binding due to a reduction in binding capacity; elevated basal 2-DG transport; and an absence of insulin stimulation of 2-DG transport. Exposure to increasing concentrations of glucose for 18 h led to a glucose concentration-dependent increase in specific insulin binding. Additionally, the various changes in the glucose-starved group were reversed after as little as 6 h of glucose refeeding. The results indicate that basal sugar transport, and insulin binding and action can be regulated by the amount of glucose in the medium.