Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Defining viability in mammalian cell cultures


A large number of assays are available to monitor viability in mammalian cell cultures with most defining loss of viability as a loss of plasma membrane integrity, a characteristic of necrotic cell death. However, the majority of cultured cells die by apoptosis and early apoptotic cells, although non-viable, maintain an intact plasma membrane and are thus ignored. Here we measure the viability of cultures of a number of common mammalian cell lines by assays that measure membrane integrity (a measure of necrotic cell death) and assays that measure apoptotic cells, and show that discrepancies in the measurement of culture viability have a significant impact on the calculation of cell culture parameters and lead to skewed experimental data.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3


  1. Goldman MH, James DC, Ison AP, Bull AT (1997) Monitoring proteolysis of recombinant human interferon-gamma during batch culture of Chinese hamster ovary cells. Cytotechnology 23:103–111

  2. Gramer MJ, Goochee CF (1993) Glycosidase activities in Chinese-hamster ovary cell lysate and cell-culture supernatant. Biotech Prog 9:366–373

  3. Gregory CD, Pound JD, Devitt A, Wilson-Jones M, Ray P, Murray RJ (2009) Inhibitory effects of persistent apoptotic cells on monoclonal antibody production in vitro. Simple removal of non-viable cells improves antibody productivity by hybridoma cells in culture. mAbs 1:370–376

  4. Hansen K, Kjalke M, Rasmussen PB, Kongerslev L, Ezban M (1997) Proteolytic cleavage of recombinant two-chain factor VIII during cell culture production is mediated by protease(s) from lysed cells—the use of pulse labelling directly in production medium. Cytotechnology 24:227–234

  5. Simpson NH, Milner AE, Al-Rubeai M (1997) Prevention of hybridoma cell death by bcl-2 during suboptimal culture conditions. Biotech Bioeng 54:1–16

  6. Singh RP, Al-Rubeai M, Gregory CD, Emery AN (1994) Cell death in bioreactors—a role for apoptosis. Biotech Bioeng 44:720–726

  7. Tey BT, Al Rubeai M (2004) Suppression of apoptosis in perfusion culture of myeloma NS0 cells enhances cell growth but reduces antibody productivity. Apoptosis 9:843–852

  8. Vermes I, Haanen C, Steffensnakken H, Reutelingsperger C (1995) A novel assay for apoptosis—flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein-labeled annexin-V. J Immunol Meth 184:39–51

Download references

Author information

Correspondence to Mohamed Al-Rubeai.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Browne, S.M., Al-Rubeai, M. Defining viability in mammalian cell cultures. Biotechnol Lett 33, 1745–1749 (2011). https://doi.org/10.1007/s10529-011-0644-2

Download citation


  • Apoptosis
  • Biopharmaceuticals
  • Mammalian cell culture
  • Necrosis
  • Viability