Abstract
To determine the influence of the inoculum age on the kinetics of hybridoma growth and metabolism, spinner flasks have been inoculated with cells previously propagated in T flasks for 43, 52, 62 and 71 hr respectively. Increasing the age of the inoculum is found to result in a longer lag phase, in a lower maximum specific growth rate and in a reduced maximal cell density. During the growth phase specific rates of glucose and glutamine uptake and of ammonia and lactate production are similar. However, with the older inoculum, much higher metabolic activities are observed during the lag phase. The production of antibodies is delayed with increasing inoculum age, but the final antibody concentrations are similar, which indicates a higher specific antibody production rate when inoculating with older cells.
Similar content being viewed by others
References
DaliliM and OllisDF (1989). Transient kinetics of hybridoma growth and monoclonal antibody production in serum-limited cultures. Biotechnol. Bioeng. 33: 984–990.
DodgeTC, JiGY and HuWS (1987). Loss of viability in hybridoma cell culture. A kinetic study. Enzyme Microb. Technol. 9: 607–611.
GeaugeyV, DuvalD, GeahelI, MarcA and EngasserJM (1989). Influence of amino acids on hybridoma cell viability and antibody secretion. Cytotechnology 2: 119–129.
GoergenJL, MartialA, MarcA and EngasserJM (1990). A kinetic model for the influence of serum in batch and continuous hybridoma cultures. In: Production of Biologicals from Animal Cells in Culture. Research, development and Achievements. Butterworths and Co. (Publishers) Ltd, Essex (in press).
GriffithsJB (1986) In: FreshneyR.I. (Ed.) Animal cell culture. IRL Press, Oxford pp. 33–69.
LevintowL and EagleH (1962). The technique of mammalian cell culture. Methods In Enzymology 5: 77–89.
MillerWM, BlanchHW and WilkeCR (1988). A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: Effect of nutrient concentration, dilution rate and pH. Biotechnol. Bioeng. 32: 947–965.
MillerWM, WilkeCR and BlanchHW (1987). Effects of dissolved oxygen concentration on hybridoma growth and metabolism in continuous culture. J. Cell. Physiol. 132: 524–530.
ReuvenyS, VelezD, MacmillanJD and MillerL (1986). Factors affecting cell growth and monoclonal antibody production in stirred reactors. J. Immunol. Meth. 86: 53–59.
ZwernerRK, CoxRM, LynnJD and ActonRT (1981). Five-year perspective of the large scale growth of mammalian cells in suspension culture. Biotechnol. Bioeng. 23: 2717–2735.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Martial, A., Dardenne, M., Engasser, JM. et al. Influence of inoculum age on hybridoma culture kinetics. Cytotechnology 5, 165–171 (1991). https://doi.org/10.1007/BF00365433
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00365433