Lethal events during gas sparging in animal cell culture

Abstract

The lethal effects of gas sparging on hybridoma cells obtained from a chemostat culture were examined in a bubble column. Experiments were performed to identify and quantify the main hazardous event: bubble formation, bubble rising, or bubble breakup. The results indicate that bubble breakup is the main cause of cell death. The protective activity of the surfactant Pluronic F68 against sparging seems to result from a direct interaction with the cells rather than influencing bubble-liquid interface properties.

Reactor Engineering in Large Scale Animal Cell Culture

“…Similar effects were observed in stirred bioreactors by sparging at different places relative to the impeller, thus giving a range of bubble sizes under identical aeration and agitation conditions with smaller bubbles again causing more damage (Oh et al 1992). Particularly clever was the way that Orton and Wang (1990) showed how cell death rates (Jö bses et al 1991;Tramper et al 1987) and liquid carryover rates (Newitt et al 1954) were similarly related to bubble size. The literature reviewed above points very clearly to bursting bubbles as the most damaging of the possible mechanism leading to loss of cell viability and total cell numbers in animal cell bioreactors.…”

“…8. Many workers also found experimentally that smaller bubbles were more damaging to cells than larger ones at the same volumetric air flow rate in bubble columns using different spargers (Tramper et al 1987;HandaCorrigan et al 1989;Jö bses et al 1991). Similar effects were observed in stirred bioreactors by sparging at different places relative to the impeller, thus giving a range of bubble sizes under identical aeration and agitation conditions with smaller bubbles again causing more damage (Oh et al 1992).…”

“…Work in bubble columns first indicated that damage associated with rising bubbles alone was not the cause as introducing air at greater depths did not cause more damage whilst high aspect ratios caused less (Handa-Corrigan et al 1989;Jö bses et al 1991;Kioukia et al 1992). Most workers therefore concluded that damage occurred due to bursting bubbles, though distinguishing it from potential damage at the sparger or in the impeller region is not easy.…”

Transfection of Spodoptera frugiperda Insect Cell and Production of Recombinant Protein

“…Kayma geriliminin böcek hücre hatlarında hücre ölümlerine sebep olmaması için tavsiye edilen havalandırma hızı 0.005-0.07 vvm karıştırma hızı ise 50-80 rpm olarak verilmiştir [15], [28]- [31].…”

CFD for Characterizing Standard and Single-use Stirred Cell Culture Bioreactors

“…Because most agitated bioreactors used in cell culture operate at relatively low mechanical power input, mechanical stress caused by bubbles can exceed that of agitation. Bubble rupture at the liquid surface was found to have the highest effect due to the high local energy density occurring when bubbles burst [Jöbses et al (1991); Chisti (2000)]. …”