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Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes

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Abstract

This study proposes an easy to use in situ device, based on multi-frequency permittivity measurements, to monitor the growth and death of attached Vero cells cultivated on microporous microcarriers, without any cell sampling. Vero cell densities were on-line quantified up to 106 cell mL−1. Some parameters which could potentially impact Vero cell morphological and physiological states were assessed through different culture operating conditions, such as media formulation or medium feed-harvest during cell growth phase. A new method of in situ cell death detection with dielectric spectroscopy was also successfully implemented. Thus, through permittivity frequency scanning, major rises of the apoptotic cell population in bioreactor cultures were detected by monitoring the characteristic frequency of the cell population, fc, which is one of the culture dielectric parameters. Both cell density quantification and cell apoptosis detection are strategic information in cell-based production processes as they are involved in major events of the process, such as scale-up or choice of the viral infection conditions. This new application of dielectric spectroscopy to adherent cell culture processes makes it a very promising tool for risk-mitigation strategy in industrial processes. Therefore, our results contribute to the development of Process Analytical Technology in cell-based industrial processes.

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Acknowledgments

The authors would like to acknowledge Sven Ansorge for his critical review of the manuscript.

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  1. Emma Petiot

    Present address: Animal Cell Technology Group, Biotechnology Research Institute, 6100 Royalmount Avenue, Montreal, H4P 2R2, QC, Canada

Authors and Affiliations

  1. Laboratoire Réactions et Génie des Procédés, UPR CNRS 3349, Nancy-Université, 2 avenue de la Forêt de Haye, 54505, Vandoeuvre-lès-Nancy Cedex, France

    Emma Petiot, Amal El-Wajgali & Annie Marc

  2. FOGALE Nanotech, 125 rue de l’Hostelerie, Ville Active Bâtiment A, Parc Acti plus, 30900, Nîmes, France

    Geoffrey Esteban

  3. Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280, Marcy L’Etoile, France

    Cécile Gény & Hervé Pinton

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  1. Emma Petiot
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  2. Amal El-Wajgali
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Correspondence to Emma Petiot.

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Petiot, E., El-Wajgali, A., Esteban, G. et al. Real-time monitoring of adherent Vero cell density and apoptosis in bioreactor processes. Cytotechnology 64, 429–441 (2012). https://doi.org/10.1007/s10616-011-9421-2

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  • DOI: https://doi.org/10.1007/s10616-011-9421-2

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