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In-situ cell density monitoring and apoptosis detection in adherent Vero cell bioreactor cultures

Authors
Journal
BMC Proceedings
1753-6561
Publisher
Springer (Biomed Central Ltd.)
Publication Date
Volume
5
Identifiers
DOI: 10.1186/1753-6561-5-s8-p8
Keywords
  • Meeting Abstract

Abstract

In-situ cell density monitoring and apoptosis detection in adherent Vero cell bioreactor cultures MEETING ABSTRACT Open Access In-situ cell density monitoring and apoptosis detection in adherent Vero cell bioreactor cultures Emma Petiot1*, Amal El-Wajgali1, Geoffrey Esteban2, Cécile Gény3, Hervé Pinton3, Annie Marc1 From 22nd European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies Vienna, Austria. 15-18 May 2011 Background In cell-based processes, and particularly in viral vaccine production, cell growth and death are strategic informa- tions to obtain for process monitoring (ie. scale-up, determination of MOI, TOI, and harvest time). Dielec- tric spectroscopy is a tool which was increasingly imple- mented on cell-culture bioreactors as it presents great potentials, compared to other methods, for the in-line monitoring of these two crucial parameters. Considering viral vaccine production, Vero cells are one of the most employed cell platform. But, due to its adherent charac- teristics, few in-line techniques were developed for cell density monitoring, on the contrary to the ones available for suspension cells. In addition, it should be underline that no in-line technique exists for quantification or detection of the mammalian cell death, despite the importance of this parameter for cell culture processes. Materials and methods The Vero cell line employed in this study was provided by Sanofi Pasteur. Cells were cultivated in serum-free conditions; either in a reference medium or in a modi- fied medium with alanine-glutamine peptide (Gluta- max®) substituted to glutamine. The adhered cell population was numbered on haemacytometer after crystal violet treatment. The apoptotic cells, labelled with annexin V, were quantified by flow cytometry (Guava). The in-line recording of permittivities at differ- ent frequencies and of the characteristic frequency of the cell population, fc, were performed by a Fogale Bio- mass system®. Theoretical background Th

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