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Pilot-scale verification of maximum tolerable hydrodynamic stress for mammalian cell culture

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Abstract

Although several scaling bioreactor models of mammalian cell cultures are suggested and described in the literature, they mostly lack a significant validation at pilot or manufacturing scale. The aim of this study is to validate an oscillating hydrodynamic stress loop system developed earlier by our group for the evaluation of the maximum operating range for stirring, based on a maximum tolerable hydrodynamic stress. A 300-L pilot-scale bioreactor for cultivation of a Sp2/0 cell line was used for this purpose. Prior to cultivations, a stress-sensitive particulate system was applied to determine the stress values generated by stirring and sparging. Pilot-scale data, collected from 7- to 28-Pa maximum stress conditions, were compared with data from classical 3-L cultivations and cultivations from the oscillating stress loop system. Results for the growth behavior, analyzed metabolites, productivity, and product quality showed a dependency on the different environmental stress conditions but not on reactor size. Pilot-scale conditions were very similar to those generated in the oscillating stress loop model confirming its predictive capability, including conditions at the edge of failure.

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Acknowledgments

The authors would like to acknowledge SNF (200020_147137/1) for financial support of this project. Furthermore, Miroslav Soos was partially supported by the Specific University Research grant of UCT (grant number 20/2015).

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Authors and Affiliations

  1. Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland

    Benjamin Neunstoecklin, Thomas K. Villiger, Massimo Morbidelli & Miroslav Soos

  2. Merck Serono S.A., Biotech Process Sciences, Corsier-sur-Vevey, Switzerland

    Eric Lucas, Matthieu Stettler & Hervé Broly

  3. Department of Chemical Engineering, University of Chemistry and Technology, Technicka 3, 16628, Prague, Czech Republic

    Miroslav Soos

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  1. Benjamin Neunstoecklin
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  2. Thomas K. Villiger
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  3. Eric Lucas
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  4. Matthieu Stettler
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  5. Hervé Broly
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  6. Massimo Morbidelli
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  7. Miroslav Soos
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Corresponding authors

Correspondence to Massimo Morbidelli or Miroslav Soos.

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Funding

This study was funded by SNF (grant number 200020_147137/1).

This study was partially funded by the Research grant of UCT (grant number 20/2015).

Conflict of interest

Benjamin Neunstoecklin declares that he has no conflict of interest.

Thomas K. Villiger declares that he has no conflict of interest.

Eric Lucas declares that he has no conflict of interest.

Matthieu Stettler declares that he has no conflict of interest.

Hervé Broly declares that he has no conflict of interest.

Massimo Morbidelli declares that he has no conflict of interest.

Miroslav Soos declares that he has no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Neunstoecklin, B., Villiger, T.K., Lucas, E. et al. Pilot-scale verification of maximum tolerable hydrodynamic stress for mammalian cell culture. Appl Microbiol Biotechnol 100, 3489–3498 (2016). https://doi.org/10.1007/s00253-015-7193-x

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