Applied Biochemistry and Biotechnology

, Volume 160, Issue 6, pp 1600–1615

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

Authors

  • Emma Petiot
    • Laboratoire des Sciences du Génie Chimique, UPR CNRS 6811Nancy-Université
  • Frantz Fournier
    • Laboratoire des Sciences du Génie Chimique, UPR CNRS 6811Nancy-Université
  • Cécile Gény
    • Sanofi pasteur
  • Hervé Pinton
    • Sanofi pasteur
    • Laboratoire des Sciences du Génie Chimique, UPR CNRS 6811Nancy-Université
Article

DOI: 10.1007/s12010-009-8674-0

Cite this article as:
Petiot, E., Fournier, F., Gény, C. et al. Appl Biochem Biotechnol (2010) 160: 1600. doi:10.1007/s12010-009-8674-0

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.

Keywords

Adherent Vero cellsSerum-free mediumSmall-scale screeningNon-invasive cell countingMicrocarriers

Nomenclature

ax

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

C

Cell concentration (cell ml−1)

DoE

Design of experiments

MR

Reference medium

MC

Complete medium

Mx

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

Gx

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

WS

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

Copyright information

© Humana Press 2009