Monitoring Growth and Death of Vero Cells Cultivated in Bioreactor with Serum-Containing and Serum-Free Media

Quesney, Sébastien; Marvel, Jacqueline; Marc, Annie; Gerdil, Catherine; Meignier, Bernard

doi:10.1007/978-94-010-0369-8_48

Cited by 10 publications

(12 citation statements)

References 4 publications

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“…The residual percentage of cells in these phases may be indicative of cell culture turnover. Similar results were found by Quesney et al in cell cycle studies with Vero cells [35]. On the other hand, and previously reported [6,18], a lower proliferation rate (represented by S and G 2 /M fractions) was found on cells cultured on PCL films at short culture times.…”

Section: Resultssupporting

confidence: 90%

Transitory oxidative stress in L929 fibroblasts cultured on poly(ε-caprolactone) films

et al. 2005

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Section: Resultssupporting

confidence: 90%

Transitory oxidative stress in L929 fibroblasts cultured on poly(ε-caprolactone) films

et al. 2005

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“…However, lysed cells were kept almost constant during the whole culture time with a value between 10% and 15% of the total cells. These results have to be compared with those obtained by Quesney et al (2001). Indeed, a more drastic cell lysis of Vero cells cultivated in serum-free media, reaching 25% of the total cells, was observed.…”

Section: Resultsmentioning

confidence: 59%

“…As previously reported, Vero cells have been recently adapted to grow in a serum-free medium for vaccine production (Butler et al, 2000;Merten et al, 1997;Petiot et al, 2010;Quesney et al, 2001;Rourou et al, 2007), but Vero cell metabolism remains not well characterized particularly when cultivated in a serum-free medium (Quesney et al, 2003). Since this cell line is of importance for virus production, a metabolic characterization has still to be established.…”

Section: Introductionmentioning

confidence: 92%

Kinetic characterization of vero cell metabolism in a serum‐free batch culture process

Petiot

Guédon

Blanchard

et al. 2010

Biotech & Bioengineering

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A global kinetic study of the central metabolism of Vero cells cultivated in a serum-free medium is proposed in the present work. Central metabolism including glycolysis, glutaminolysis, and tricarboxylic acid cycle (TCA) was demonstrated to be saturated by high flow rates of consumption of the two major substrates, glucose, and glutamine. Saturation was reavealed by an accumulation of metabolic intermediates and amino acids, by a high production of lactate needed to balance the redox pathway, and by a low participation of the carbon flow to the TCA cycle supply. Different culture conditions were set up to reduce the central metabolism saturation and to better balance the metabolic flow rates between lactate production and energetic pathways. From these culture conditions, substitutions of glutamine by other carbon sources, which have lower transport rates such as asparagine, or pyruvate in order to shunt the glycolysis pathway, were successful to better balance the central metabolism. As a result, an increase of the cell growth with a concomitant decrease of cell death and a better distribution of the carbon flow between TCA cycle and lactate production occurred. We also demonstrated that glutamine was a major carbon source to supply the TCA cycle in Vero cells and that a reduction of lactate production did not necessary improve the efficiency of the Vero cell metabolism. Thus, to adapt the formulation of the medium to the Vero cell needs, it is important to provide carbon substrates inducing a regulated supply of carbon in the TCA cycle either through the glycolysis or through other pathways such as glutaminolysis. Finally, this study allowed to better understand the Vero cell behavior in serum-free medium which is a valuable help for the implementation of this cell line in serum-free industrial production processes.

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“…On one side, cell and virus propagation were investigated in different cultivation systems (static systems, roller bottles, spinner, stirred tank). On the other side, not only different media (serum-containing (SC) as well as serum-free (SF)) but also different microcarriers (Cytodex 1 and 3) were used for large-scale cultures (Merten et al 1996;Quesney et al 2001;Kistner et al 2007;Pereira 1998, 1995). Additional variations resulted from the use of MDCK as well as Vero cell lines from different sources and the use of different influenza virus strains ).…”

Section: Introductionmentioning

confidence: 99%

MDCK and Vero cells for influenza virus vaccine production: a one-to-one comparison up to lab-scale bioreactor cultivation

Genzel

Dietzsch

Rapp

et al. 2010

Appl Microbiol Biotechnol

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Over the last decade, adherent MDCK (Madin Darby canine kidney) and Vero cells have attracted considerable attention for production of cell culture-derived influenza vaccines. While numerous publications deal with the design and the optimization of corresponding upstream processes, one-to-one comparisons of these cell lines under comparable cultivation conditions have largely been neglected. Therefore, a direct comparison of influenza virus production with adherent MDCK and Vero cells in T-flasks, roller bottles, and lab-scale bioreactors was performed in this study. First, virus seeds had to be adapted to Vero cells by multiple passages. Glycan analysis of the hemagglutinin (HA) protein showed that for influenza A/PR/8/34 H1N1, three passages were sufficient to achieve a stable new N-glycan fingerprint, higher yields, and a faster increase to maximum HA titers. Compared to MDCK cells, virus production in serum-free medium with Vero cells was highly sensitive to trypsin concentration. Virus stability at 37 degrees C for different virus strains showed differences depending on medium, virus strain, and cell line. After careful adjustment of corresponding parameters, comparable productivity was obtained with both host cell lines in small-scale cultivation systems. However, using these cultivation conditions in lab-scale bioreactors (stirred tank, wave bioreactor) resulted in lower productivities for Vero cells.

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Monitoring Growth and Death of Vero Cells Cultivated in Bioreactor with Serum-Containing and Serum-Free Media

Cited by 10 publications

References 4 publications

Transitory oxidative stress in L929 fibroblasts cultured on poly(ε-caprolactone) films

Transitory oxidative stress in L929 fibroblasts cultured on poly(ε-caprolactone) films

Kinetic characterization of vero cell metabolism in a serum‐free batch culture process

MDCK and Vero cells for influenza virus vaccine production: a one-to-one comparison up to lab-scale bioreactor cultivation

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