Human lymphoblastoid cell line WI-L2–729-HFZ was fused with human lymphnode lymphocytes in one fusion and with human spleen cells in another fusion to generate human-human hybridomas. In both, increasing doses of insulin were added to the HAT medium immediately after the PEG-mediated cell fusion (10-1-10-5 IU/ml) and the number of clones formed was determined 3 weeks later. 10-3 IU/ml of insulin resulted in a 2- to 5-fold increase in the number of clones generated compared to the control plates. In view of the known difficulties in generating high-yield human-human hybridoma fusions, it is suggested that the use of insulin-supplemented HAT medium may provide a more efficient way in obtaining such clones.

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