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Characterization of Vero cell growth and death in bioreactor with serum-containing and serum-free media

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Abstract

The density of viable cells in a culture results from a balance between cell proliferation and cell death. The aim of this study was to characterize and compare these two phenomena in Vero cell cultures in one serum containing medium (ScA) and one serum free medium (SfB) in bioreactors. Cell growth was evaluated by cell counting(after crystal violet staining) and cell cycle analysis. Necrosis and apoptosis were characterized and quantified by measuring the release of LDH, trypan blue exclusion,annex in V-FITC/PI staining and TUNEL assay. ScA supported a higher maximal viable-cell density(2.3 × 106 vs. 1.8 × 106 cells ml-1). However, cell cycle analysis showed that cell division was more active in SfB than in ScA. LDH release in the supernatant increased much earlier in SfB than in ScA (one vs. five days), but trypan blue counts showed no apparent difference in the viability of the cultures. Apoptosis, evidenced by annexin V-FITC/PI staining, could be detected in the population of suspension cells detached from microcarriers, but not among adherent cells; positivity of the TUNEL assay occurred later than that of the annexin V-FITC/PI staining. Our data indicate that the lower cell yield in SfB,compared with that in ScA, results from a higher cell death rate. Apparently, cells die from apoptosis followed by secondary necrosis.

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Authors and Affiliations

  1. Development Department, Aventis Pasteur, Campus Mérieux, 1541 Avenue Marcel Mérieux, F-69280, Marcy L'étoile, France

    Sébastien Quesney & Bernard Meignier

  2. Centre Européen de Recherche en Virologie et Immunologie, INSERM, unité 503, 21 Avenue Tony Garnier, F-69365, Lyon, France

    Jacqueline Marvel

  3. Laboratoire des Sciences du Génie Chimique, CNRS, UPR 6811, ENSIC, 1 Rue Granville, F-54001, Nancy, France

    Annie Marc

  4. Development Department, Aventis Pasteur, Campus Mérieux, 1541 Avenue Marcel Mérieux, F-69280, Marcy L'étoile, France

    Catherine Gerdil

Authors
  1. Sébastien Quesney
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  2. Jacqueline Marvel
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  3. Annie Marc
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  4. Catherine Gerdil
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  5. Bernard Meignier
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Correspondence to Catherine Gerdil.

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Quesney, S., Marvel, J., Marc, A. et al. Characterization of Vero cell growth and death in bioreactor with serum-containing and serum-free media. Cytotechnology 35, 115–125 (2001). https://doi.org/10.1023/A:1017589526145

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