Evidence against a role for insulin-like growth factor II in the autonomous growth of rat 18,54-SF cells.

The 18,54-SF rat cell line multiplies in serum-free medium and has been reported to produce insulin-like growth factor II (IGF-II) and to possess IGF-II/mannose 6-phosphate (Man-6-P) receptors, raising the possibility of autocrine growth stimulation by IGF-II acting through this receptor. When serum-free medium was changed every 24 h the 18,54-SF cells multiplied at the same rate as when the medium was not changed. An antibody (No. 3637) which blocks the binding of IGF-II to the IGF-II/Man-6-P receptor did not decrease or increase the growth rate of the 18,54-SF cells when medium was changed every 24 h. When the medium was changed every 12 h the rate of accumulation of cells in the culture was decreased. Addition of IGF-I or IGF-II at 10, 50, and 100 ng/ml every 12 h did not correct this decrease in cell number. When the medium was replaced every 12 h, cells in the periphery of the cultures gradually became nonviable as assessed by trypan blue uptake. However, the percentage of cells synthesizing DNA in the center of the cultures (approximately 50% during a 3 h pulse with tritiated thymidine) was the same whether the medium was changed every 12 h or left unchanged. Addition of IGF-I or IGF-II to the fresh medium change every 12 h did not increase the percentage of cells synthesizing DNA.