The overall objective of this proposal is to characterize the morphology, structure, geometry, and rheological properties of two representative hybridoma cell lines in order to evaluate the effects of well defined hydrodynamic forces on such cells, and to establish membrane failure criteria that result in cell damage and cell death. A range of experimental techniques (electron, light and fluorescence microscopy, micropipet manipulation, rheoscopy, and micro-rheometry) will be used to evaluate the aforementioned properties, and to determine the influence of hydrodynamic forces on the cells in simple (laminar and turbulent) flow, and in complex turbulent flows that model more closely the fluid environment in bioreactors. Mechanisms of membrane and cell damage will be evaluated by combining experimental measurements with fluid flow studies and theoretical calculations. These studies are essential to improve and/or simplifiy bioreactor design, and to promote the optimal growth conditions, for the mass production of monoclonal antibodies.
