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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

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