Previous in vivo studies have shown that growth hormone (GH) affects vitamin D and mineral metabolism. Insulin-like growth factor-I (IGF-I) was recently reported to be a regulator of renal 1,25-dihydroxyvitamin D 3 (1,25-(OH) 2 D 3 ) production, suggesting that it mediates the effects of GH on vitamin D metabolism. However, there is no direct evidence to support this. The present study was designed to investigate the in vitro effects of GH and IGF-I on the renal production of 1,25- (OH) produced were determined after lipid extraction and HPLC purification. Production of 1,25-(OH) 2 D 3 and 24,25-(OH) 2 D 3 was increased after both IGF-I and GH preincubation in a dose-dependent manner. Significant increases were found after preincubation with 13 nmol/l IGF-I (1,25-(OH) 2 D 3 , 1.8-fold; 24,25-(OH) 2 D 3 , 1.5-fold) or 0.9 or 9 nmol/l GH (1,25-(OH) 2 D 3 , 1.3-fold and 1.5-fold; 24,25-(OH) 2 D 3 , 1.4-fold and 1.5-fold respectively). Furthermore, the effect of 9 nmol/l GH on 1,25-(OH) 2 D 3 and 24,25-(OH) 2 D 3 production was blocked in the presence of IGF-I receptor monoclonal antibody. These results confirm that IGF-I acts on renal tubules, resulting in induction of 1,25-(OH) 2 D 3 and 24,25-(OH) 2 D 3 production, and the findings suggest that GH stimulates 1,25-(OH) 2 D 3 and 24,25-(OH) 2 D 3 production by increasing local IGF-I production in the kidney.