Influence of dissolved oxygen concentration on growth, mitochondrial function and antibody production of hybridoma cells in batch culture

Abstract

The effects of dissolved oxygen concentration on hybridoma cell growth, metabolism, and antibody production were studied. A mouse hybridoma cell line producing an IgG1 directed at a consensus α-interferon was grown in batch cultures in a 5 dm3 stirred bioreactor at dissolved oxygen (DO) concentrations of 5, 30, 90 and 95% or air saturation. High oxygen tension (95% of air saturation) reduced specific growth rate without affecting cell viability. At lower dissolved oxygen levels, specific growth rates were approximately independent of DO, although changes in mitochondrial function and antibody production were observed. Flow cytometry assays of a fluorescent mitochondria-specific marker (Rhodamine 123) show significant single-cell heterogeneities during late exponential growth and greater average fluorescence in cultures grown at 95% DO. The quantity of cell-surface immunoglobulin, measured by an immunofluorescent flow cytometric technique, was the same at high (95%) and low (5%) dissolved oxygen concentrations. Myeloma cells of the type used in constructing the above hybridoma line were much less sensitive to dissolved oxygen level. Specific respiration rate, pyruvate utilization rate, cytochrome oxidase activity, and succinate-cytochrome c oxidoreductase activity were significantly greater (62–116%) for the hybridoma cells than for the myeloma cells in T-flask cultures.