Modeling industrial centrifugation of mammalian cell culture using a capillary based scale‐down system

Westoby, Matthew; Rogers, Jameson K.; Haverstock, Ryan; Romero, Jonathan K.; Pieracci, John

doi:10.1002/bit.23051

Cited by 32 publications

(36 citation statements)

References 23 publications

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“…Continuing a quantitative approach to understanding the effect of hydrodynamic forces in downstream processing equipment on animal cells, Westoby et al (2011) used EDR measurements in a capillary shear device to model industrial scale continuous-flow centrifugation. They reported that exposing cells to a EDR value of 3 9 10 4 W/kg (3 9 10 7 W/m 3 ) was a critical EDR value at which they could mimic the compromised performance of the large scale centrifuges.…”

Section: Cell Damage In Downstream Equipment and Other Bioprocess Equmentioning

confidence: 99%

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

Hu¹,

2011

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Suspension animal cell culture is now routinely scaled up to bioreactors on the order of 10,000 L, and greater, to meet commercial demand. However, the concern of the 'shear sensitivity' of animal cells still remains, not only within the bioreactor, but also in the downstream processing. As the productivities continue to increase, titer of ~10 g/L are now reported with cell densities greater than 2 × 10(7) cells/mL. Such high, and potentially higher cell densities will inevitably translate to increased demand in mass transfer and mixing. In addition, achieving productivity gains in both the upstream stage and downstream processes can subject the cells to aggressive environments such as those involving hydrodynamic stresses. The perception of 'shear sensitivity' has historically put an arbitrary upper limit on agitation and aeration in bioreactor operation; however, as cell densities and productivities continue to increase, mass transfer requirements can exceed those imposed by these arbitrary low limits. Therefore, a better understanding of how animal cells, used to produce therapeutic products, respond to hydrodynamic forces in both qualitative and quantitative ways will allow an experimentally based, higher, "upper limit" to be created to guide the design and operation of future commercial, large scale bioreactors. With respect to downstream hydrodynamic conditions, situations have already been achieved in which practical limits with respect to hydrodynamic forces have been experienced. This review mainly focuses on publications from both the academy and industry regarding the effect of hydrodynamic forces on industrially relevant animal cells, and not on the actual scale-up of bioreactors. A summary of implications and remaining challenges will also be presented.

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Section: Cell Damage In Downstream Equipment and Other Bioprocess Equmentioning

confidence: 99%

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

Hu¹,

2011

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“…As these tools continue to evolve, integration of primary recovery is being seen as increasingly important in the development process, to understand its impact on the release of process related impurities. This is being achieved by the development of a number of scale‐down methodologies for these process operations, including for centrifugation, microfiltration, and depth filtration …”

Section: Introductionmentioning

confidence: 99%

Differential response in downstream processing of CHO cells grown under mild hypothermic conditions

Tait

Tarrant

Velez-Suberbie

et al. 2013

Biotechnology Progress

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The manufacture of complex therapeutic proteins using mammalian cells is well established, with several strategies developed to improve productivity. The application of sustained mild hypothermic conditions during culture has been associated with increases in product titer and improved product quality. However, despite associated cell physiological effects, very few studies have investigated the impact on downstream processing (DSP). Characterization of cells grown under mild hypothermic conditions demonstrated that the stationary phase was prolonged by delaying the onset of apoptosis. This enabled cells to maintain viability for extended periods and increase volumetric productivity from 0.74 to 1.02 g L−1. However, host cell proteins, measured by ELISA, increased by ∼50%, attributed to the extended time course and higher peak and harvest cell densities. The individual components making up this impurity, as determined by SELDI-TOF MS and 2D-PAGE, were shown to be largely comparable. Under mild hypothermic conditions, cells were less shear sensitive than those maintained at 37°C, enhancing the preliminary primary recovery step. Adaptive changes in membrane fluidity were further investigated by adopting a pronounced temperature shift immediately prior to primary recovery and the improvement observed suggests that such a strategy may be implementable when shear sensitivity is of concern. Early and late apoptotic cells were particularly susceptible to shear, at either temperature, even under the lowest shear rate investigated. These findings demonstrate the importance of considering the impact of cell culture strategies and cell physiology on DSP, by implementing a range of experimental methods for process characterization. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:688–696, 2013

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“…This data demonstrates that after centrifugation, an increase in the number of cells present in smaller aggregate sizes occurred. This would imply that the centrifugation process step may have reduced the larger aggregates of cells into smaller aggregates or into single cells, which is often reported with mammalian cells [42]. However, the lack of a greater number of cells in larger aggregates in the pre-centrifugation samples may not support this conclusion.…”

Section: Live Cell Aggregate Sizementioning

confidence: 77%

Comparability of scalable, automated hMSC culture using manual and automated process steps

Archibald

Chandra

Thomas

et al. 2016

Biochemical Engineering Journal

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a b s t r a c tAutomation will likely to play a key role in the development of scalable manufacturing processes for cell-based therapies. In this study, we have compared the effects of manual centrifugation and automated non-centrifugation cell culture process steps, performed using TAP biosystems' CompacT SelecT automated cell culture platform, upon hMSC morphology, number, viability, surface marker expression, Short tandem repeat (STR) profile, and paracrine function. Furthermore, the comparability between flow cytometry analyses of hMSCs, performed at multiple sites, was investigated. No significant difference in hMSC growth and characteristics was observed between cells cultured using either the manual centrifugation process step or the automated non-centrifugation process step, in which residual dissociation agent is carried over. However, some variability in paracrine activity was observed between hMSCs cultured using alternative process steps. It is also apparent that differences in analytical methods can influence the inter-laboratory reproducibility of hMSC flow cytometry analysis, although differences in culture may also contribute to the variability observed in the expression of 2 of the 8 surface markers examined. This novel investigation into the effects of these two key process steps will help to improve the understanding of the influence of automated cell culture upon various cell culture parameters, as well as upon process comparability.

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Modeling industrial centrifugation of mammalian cell culture using a capillary based scale‐down system

Cited by 32 publications

References 23 publications

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

The potential of hydrodynamic damage to animal cells of industrial relevance: current understanding

Differential response in downstream processing of CHO cells grown under mild hypothermic conditions

Comparability of scalable, automated hMSC culture using manual and automated process steps

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