title: Pilot-scale verification of maximum tolerable hydrodynamic stress for mammalian cell culture.
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.
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Neunstoecklin
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland.
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Villiger
authors.affiliation: Merck Serono S.A., Biotech Process Sciences, Corsier-sur-Vevey, Switzerland.
authors.forename: Eric
authors.initials: E
authors.lastname: Lucas
authors.affiliation: Merck Serono S.A., Biotech Process Sciences, Corsier-sur-Vevey, Switzerland.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Stettler
authors.affiliation: Merck Serono S.A., Biotech Process Sciences, Corsier-sur-Vevey, Switzerland.
authors.forename: Hervé
authors.initials: H
authors.lastname: Broly
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland. massimo.morbidelli@chem.ethz.ch.
authors.forename: Massimo
authors.initials: M
authors.lastname: Morbidelli
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland. miroslav.soos@chem.ethz.ch.|Department of Chemical Engineering, University of Chemistry and Technology, Technicka 3, 16628, Prague, Czech Republic. miroslav.soos@chem.ethz.ch.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Soos
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002460:Cell Line; D049109:Cell Proliferation; D006801:Humans; D057446:Hydrodynamics; D010865:Pilot Projects
keywords: Downscaling; Mammalian cell culture; Maximum tolerable hydrodynamic stress; Pilot scale; Upscaling