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Rapid Screening of Serum-Free Media for the Growth of Adherent Vero Cells by Using a Small-Scale and Non-invasive Tool

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Abstract

The paper proposes a rapid screening method for a first step improvement of an animal component-free medium dedicated to the growth of the anchorage-dependent Vero cell line. A new, rapid, and non-invasive technique is presented to specifically monitor cultures of adherent cells in 96-well plates. The operating conditions of an image analyzer are adapted to take into account the decrease of cell size when the attached cell density increases. An experimental design is carried out to assess the influence of ten component groups in the original medium. Two groups including protein extracts, growth factor, insulin, glucose, and pyruvate show significant positive effects. The groups with vitamins and molecules related to nitrogenous bases display a less pronounced influence. The mixture of amino acids, B1 vitamin, magnesium sulfate, and sodium phosphate as well as the couple sodium citrate and ferric chloride lead to a downward trend. The screening results are proved to be scalable in stirred cultures with cells on microcarriers. An improved serum-free medium, with some component groups being removed or added, can be rapidly formulated to reach respectively similar or 1.6 times higher cell density than in the original medium. The results from this global approach could be helpful to further focus experiments on identified medium components.

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Abbreviations

a x :

Coefficients of the polynomial model (x = 1 to 10)

C :

Cell concentration (cell ml−1)

DoE:

Design of experiments

M R :

Reference medium

M C :

Complete medium

M x :

Screening media formulated for DoE (x = 1 to 16)

PA:

“Cellscreen” device module for adherent cell analysis

PS:

“Cellscreen” device module for suspension cell analysis

G x :

Groups of components present in M C medium (x = 1 to 10)

WS:

Percentage of well surface covered by Vero cells in the 96-well plates (%)

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Acknowledgment

The authors are grateful to Dr. Isabelle Chevalot (LSGC-CNRS, Nancy) for the help she provided on the use of the “Cellscreen” device.

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Authors and Affiliations

  1. Laboratoire des Sciences du Génie Chimique, UPR CNRS 6811, Nancy-Université, 2 avenue de la Forêt de Haye, 54505, Vandoeuvre-lès-Nancy Cedex, France

    Emma Petiot, Frantz Fournier & Annie Marc

  2. Sanofi pasteur, 1541 avenue Marcel Mérieux, 69280, Marcy L’Etoile, France

    Cécile Gény & Hervé Pinton

Authors
  1. Emma Petiot
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  2. Frantz Fournier
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  3. Cécile Gény
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  4. Hervé Pinton
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  5. Annie Marc
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Correspondence to Annie Marc.

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Petiot, E., Fournier, F., Gény, C. et al. Rapid Screening of Serum-Free Media for the Growth of Adherent Vero Cells by Using a Small-Scale and Non-invasive Tool. Appl Biochem Biotechnol 160, 1600–1615 (2010). https://doi.org/10.1007/s12010-009-8674-0

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  • DOI: https://doi.org/10.1007/s12010-009-8674-0

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