Enhancement of hybridoma formation by addition of insulin to HAT medium (HIAT)

The effects of insulin on the formation of hybridoma clones following fusion experiments with SRBC immunized BALB/c mouse spleen cells and P3U1 mouse plasmacytoma cells were evaluated. The addition of insulin to HAT medium (HIAT) resulted in significant increases in the number and size of hybridoma colonies generated. The total number of anti-SRBC antibody-secreting clones also increased as much as sevenfold using insulin-supplemented medium compared to HAT alone. In view of the increasing interest in hybridoma technology for monoclonal antibody production, the use of insulin-supplemented medium (HIAT) may significantly expedite ongoing work by providing a more efficient method for the establishment of stable clones.