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 media viz IMDM and RPMI with FCS (3% & 5%). The parameters such as cell count, cell viability and avidity were determined on a time course basis from day 1 to day 18, to assess the optimization of the roller bottle process for the production of anti – D and to determine the influence of factors namely medium, serum concentration and varying cell density. The cell viability and cell count remained significant upto 8 or 10 days. Cell viability assessed by dye binding revealed that apoptosis of hybridoma cells sets suddenly after 10 days in all culture bottles. The observations implied that 0.2 106/ ml seemed to be optimum for growth of hybridoma. IMDM seemed to be comparatively superior to RPMI. The study also warrants empirical informations of H9r9 cell metabolism to achieve optimization. Keywords: Hybridoma (H9r9), Foetal calf serum (FCS), Anti – D, Apoptosis, IMDM/RPMI Medium Received: 12 January 2009 / Received in revised form: 6 February 2009, Accepted: 25 February 2009, Published online: 28 February 2009

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