Determinant factors for hybridmyeloma culture (H9r9) in the yield of anti - D – impact of medium (IMDM and RPMI), serum (FCS) and cell density

The hybridoma (H9r9) were cultured in roller bottles with two mediaviz IMDM and RPMI with FCS (3% & 5%). The parameters such ascell count, cell viability and avidity were determined on a timecourse basis from day 1 to day 18, to assess the optimization of theroller bottle process for the production of anti – D and to determinethe influence of factors namely medium, serum concentration andvarying cell density. The cell viability and cell count remainedsignificant upto 8 or 10 days. Cell viability assessed by dye bindingrevealed that apoptosis of hybridoma cells sets suddenly after 10days in all culture bottles. The observations implied that 0.2 × 106 /ml seemed to be optimum for growth of hybridoma. IMDM seemedto be comparatively superior to RPMI. The study also warrantsempirical informations of H9r9 cell metabolism to achieveoptimization.