Development of a protein-free medium with ferric citrate substituting transferrin for the cultivation of mouse-mouse hybridomas.

inhibitor of the lipid peroxidation induced by iron salts and ascorbic acid.7) It has been revealed that mammalian cells in culture do not always Tequire transferrin, ifiTon is provided as a complex with appropriate chelators.8) To design a protein-free medium for hybridomas, we studied the effect of ferric citratc as an alternative to transferrin on thc proliferation of hybridomas. Mouse hybridoma IA-4-) producing IgG2u specific to akabane virus was used in this study. The hybridoma was a fusion product between the mouse myeloma cell linc, P3-X63-Ag8-Ul.iO) and BALBic spleen cells. The hybridoma was rnaintained in DF mediumii) Bupplcmented with lepg!mi of insulin (I; Nevo), 35pg!ml of human transferrin (T; Sigma), IO"M ethanolamine (E) and 2.S nM selenium (S) CITES)') at 37'C in humidified S% COi195% air, Cells werc inoculated into 35mm plastic dishes (Falcon) and cultured in DF medium supplernented with various combinations of growth factors. lron compound