title: Adaptation for survival: phenotype and transcriptome response of CHO cells to elevated stress induced by agitation and sparging.
abstract: In this work, the response and adaption of CHO cells to hydrodynamic stress in laboratory scale bioreactors originating from agitation, sparging and their combination is studied experimentally. First, the maximum hydrodynamic stress, τ(max), is characterized over a broad range of operating conditions using a shear sensitive particulate system. Separate stress regimes are determined, where τ(max) is controlled either by sparging, agitation, or their combination. Such conditions are consequently applied during cultivations of an industrial CHO cell line to determine the cellular responses to corresponding stresses. Our results suggest that the studied CHO cell line has different threshold values and response mechanisms for hydrodynamic stress resulting from agitation or sparging, respectively. For agitation, a characteristic local minimum in viability was found after stress induction followed by viability recovery, while at highest sparging stress a monotonic decrease in viability was observed. If both stresses were combined, also both characteristic stress responses could be observed, amplifying each other. On the other hand, cellular metabolism, productivity and product quality did not change significantly. Transcriptome analysis using mRNA microarrays confirmed that separate adaptation mechanisms are activated in the different stress situations studied, allowing identification of these stresses using a transcriptome fingerprinting approach. Functional analysis of the transcripts was consequently used to improve our understanding of the molecular mechanisms of shear stress response and adaptation.
authors.affiliation: Novartis Pharma AG, Biologics Process R&D, Basel, Switzerland; Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, Switzerland.
authors.forename: Jochen B
authors.initials: JB
authors.lastname: Sieck
authors.affiliation: Novartis Pharma AG, Biologics Process R&D, Basel, Switzerland.
authors.forename: Wolfgang E
authors.initials: WE
authors.lastname: Budach
authors.affiliation: Novartis Pharma AG, Biologics Process R&D, Basel, Switzerland.
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Suemeghy
authors.affiliation: Novartis Pharma AG, Biologics Process R&D, Basel, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Leist
authors.affiliation: Novartis Pharma AG, Biologics Process R&D, Basel, Switzerland.
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Villiger
authors.affiliation: Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, Switzerland.
authors.forename: Massimo
authors.initials: M
authors.lastname: Morbidelli
authors.affiliation: Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, Switzerland. Electronic address: miroslav.soos@chem.ethz.ch.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Soos
mesh_terms: D000818:Animals; D019149:Bioreactors / Q000382:microbiology*; D016466:CHO Cells; D003412:Cricetulus; D012333:RNA, Messenger; D059467:Transcriptome / Q000235:genetics*
chemical_list: D012333:RNA, Messenger
keywords: CHO cells; Environomics; Stirred bioreactors; Stress adaptation; Stress response; Transcriptome analysis; Transcriptome fingerprinting; mRNA microarrays