The effect of insulin on human-mouse hybridoma formation

To evaluate the effect of insulin on the formation of human-mouse hybridoma clones, P3U1 mouse plasmacytoma cells were fused with human lymph-node lymphocytes in the presence of polyethylene glycol. After fusion, cells were grown for 2 weeks in HAT medium supplemented with insulin (HIAT, 10^-1-10^-5 units/ml) or in HAT medium alone. The addition of 10^-3 units/ml of insulin to HAT medium resulted in an over two fold increase in the number of clones formed and in the average colony size compared to growing the fused cells in HAT medium alone. In view of the recent increasing interest in human-mouse hybridoma fusions it is suggested that selective medium HAT should be supplemented by insulin (HIAT) to enhance the number of colonies formed and provide a more efficient way for stabilizing the newly formed hybrids.