Optimal choice method of detection of the viability of cell cultures for tests on proliferation and cytotoxicity

DOI: 10.29296/2618723X-2021-02-03

A.N. Afanaseva, Jr. research assistant, ORCID: 0000-0002-1443-4294,
V.B. Saparova, Research associate, ORCID: 0000-0002-8445-1129,
T.A. Selmenskikh, Research worker, ORCID: 0000-0001-7073-311X,
I.E. Makarenko, Head of the Medical Department, ORCID: 0000-0003-2308-0608

«Pharm-Holding» ZAO,
34 A Svyazi Street, Strelna, Saint-Petersburg, 198515, Russia

E-mail: [email protected]

Keywords: cell viability MTT test formazan tetrazolium salt resazurin resorufin luminescence analysis adenositriphosphate firefly luciferase CHO-K1 32D clone 3

For citation:

Afanaseva A.N. , Saparova V.B., Selmenskikh T.A., Makarenko I.E. Optimal choice method of detection of the viability of cell cultures for tests on proliferation and cytotoxicity. Laboratory Animals for Science. 2021; 2. https://doi.org/10.29296/2618723X-2021-02-03


Viability studies are carried out to measure the proportion of viable cells after exposure to a substance, drug or manipulation, for example, in studies of cytotoxicity or proliferative activity, after cryostorage or cell division, and etc. In these studies, any in vitro changes can be interpreted to predict the in vivo response of the same or similar cells. The European Medicines Agency (EMA) supports the implementation of the so-called 3Rs principles – replace, reduce and refine – for the ethical use of animals in medicine testing. This article has been written in compliance with the 3Rs principle and does not describe any studies of animals as objects.

Currently, there are several methods for assessing the viability of cells, based on various biochemical processes taking place inside the cell. These methods can include both direct detection of ATP and determination of the activity of mitochondrial dehydrogenases. The choice of the method used in the study will depend on the agent being studied, the nature of the expected response, on the target cells, and, in particular, on the variability of the data applicable in each case.

The aim of this study was to perform a comparative analysis of three common methods for determining cell viability: MTT test, analysis of living cells using resazurin, and CellTiter-Glo® luminescence analysis.

Material and methods. Selected methods for determining viability: MTT test, analysis of living cells using resazurin, luminescence analysis of cell viability CellTiter-Glo®. As cell lines, various lines were used according to the type of cell culture growth: adherent (CHO-K1 – Chinese hamster ovary cells) and mixed, adherent and suspension (32D clone 3 – mouse bone marrow cells). Evaluation parameters were determination of the sensitive range of cell concentrations and the coefficient of variation (CV) within each concentration.

Results. According to the results of the study, the most sensitive, having low levels of variability in the concentration range and, accordingly, the most optimal, were the methods – analysis of living cells using resazurin and luminescent analysis of cell viability CellTiter-Glo®.

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Authors contributions 

A.N. Afanasyeva – significant contribution to the concept or design of the work; collecting, analyzing or interpreting the results of work, writing a text, a critical revision of its content, setting up experiments

V.B. Saparova – assistance in the analysis and interpretation of the results of the work

T.A. Selmenskikh – setting up experiments, collecting data, participating in the analysis of results

I.E. Makarenko – approval of the final version of the article for publication; agreement to be responsible for all aspects of the work, due study and resolution of issues related to the accuracy of data or the integrity of all parts of the article.

Conflict of interest

The authors declare that there is no conflict of interest.


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