Serum-free medium for hybridoma and parental myeloma cell cultivation: a novel composition of growth-supporting substances

Abstract

Serum-free chemically defined medium for hybridoma and parental myeloma cultivation was developed on the basis of testing of individual substances supporting hybridoma growth under serum-free conditions. Optimized concentrations of transferrin, insulin, ethanolamine, linoleic acid, serum albumin, ascorbic acid, hydrocortisone, and trace elements could substitute serum. Developed serum-free hybridoma (SFH) medium differs from analogous previously described media mainly by a more complete combination of growth-supporting supplements and by the presence of ascorbic acid and hydrocortisone. Growth comparable with that in the medium supplemented with 10% bovine serum was achieved with four hybridomas and two myelomas. SFH medium was also suitable for long-term cultivation of hybridomas without cessation of monoclonal antibody production. Growth potency and the specific growth requirements of hybridomas in serum-free medium are, to a large degree, determined by parental myeloma.