Cytotechnology

, Volume 35, Issue 2, pp 115–125

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

  • Sébastien Quesney
  • Jacqueline Marvel
  • Annie Marc
  • Catherine Gerdil
  • Bernard Meignier
Article

DOI: 10.1023/A:1017589526145

Cite this article as:
Quesney, S., Marvel, J., Marc, A. et al. Cytotechnology (2001) 35: 115. doi:10.1023/A:1017589526145

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.

apoptosiscell cyclelactate dehydrogenasemicrocarriersnecrosisserum-free mediumTUNELVero cells

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Sébastien Quesney
    • 1
  • Jacqueline Marvel
    • 2
  • Annie Marc
    • 3
  • Catherine Gerdil
    • 4
  • Bernard Meignier
    • 1
  1. 1.Development DepartmentAventis PasteurMarcy L'étoileFrance
  2. 2.Centre Européen de Recherche en Virologie et ImmunologieINSERMLyonFrance
  3. 3.Laboratoire des Sciences du Génie ChimiqueCNRS, UPR 6811, ENSICNancyFrance
  4. 4.Development DepartmentAventis PasteurMarcy L'étoileFrance