Polyvinyl alcohol and polyethylene glycol as protectants against fluid-mechanical injury of freely-suspended animal cells (CRL 8018)

Abstract

Two identical bioreactors run in parallel were used to examine the phenomenological characteristics of two additives, polyethylene glycol (PEG) and polyvinyl alcohol (PVA), used as protectants against fluid-mechanical cell damage. Cell-protecting ability was evaluated by comparing apparent cell growth rates of freely suspended CRL-8018 hybridoma cells cultured in serum-free medium under surface aerated conditions whereby cell damage is due to bubble entrainment and breakup. PEG of various molecular weights was used to determine whether the size of the polymer has significant effects on PEG's cell-protecting capabilities. All the PEG's with molecular weights larger than 1400 showed similar protective effects. The effect of PEG concentration was then evaluated and results showed that concentrations greater than 0.05% w/v did not significantly improve the cell-protecting properties. Direct comparisons made between the PVA, PEG, and pluronic F68 as cell protectants showed that PEG protected cells better than F68 and that PVA provided even better protection than PEG. The mechanism of protection, fluid-mechanical or biological in nature, was examined by growing the cells in additive from the beginning of the experiment (long-term exposure), or adding the additive after the cells had been agitated at rates detrimental to the cells (short-term exposure). In agreement with results reported previously on PEG and F68, fast-acting protection was seen. This implies a fluid-mechanical rather than a biological protection mechanism. In an attempt to correlate interfacial properties of the resulting media with shear protection, interfacial tension and viscosity measurements of all the media were made. On the basis of these measurements, we find no definitive correlations for evaluating these additives' cell-protecting capabilities.