The MTT Tetrazolium Salt Assay Scrutinized: How to Use this Assay Reliably to Measure Metabolie Activity of Cell Cultures in vitro for the Assessment of Growth Characteristics, IC₅₀-Values and Cell Survival

Abstract

Summary:The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay is widely used for in vitro measurement of the metabolic viability of cell cultures subjected to different culture conditions. This convenient assay, which is based on the ability of viable cells to produce formazan, can be affected significantly by a number of conditions. These conditions can be roughly divided into two groups, firstly influences which affect the spectrum of the produced formazan and secondly influences which affect the amount of formazan produced per cell. We studied the various chemical and biochemical aspects involved in the MTT assay. Our data indicate that microscopical viewing of cell cultures before and after performing the assay, a medium renewal with a well-defined MTT-incubation medium at the end of the culture period and regular cell counting are essential steps to ensure a reliable performance of the MTT assay. In conclusion, providing the necessary precautions are taken, the MTT assay can be used reliably to measure metabolic activity of cell cultures in vitro for the assessment of growth characteristics^ IC 50 -values and cell survival.

Applicability of the MTT assay for measuring viability of cyanobacteria and algae, specifically for Microcystis aeruginosa (Chroococcales, Cyanobacteria)

“…This has also been reported for other cell cultures (Sieuwerts et al 1995;Zhang & Cox 1996;Bernhard et al 2003) but has never been reported in cyanobacteria or algae. Capasso et al (2003) compared viability of nitrogenlimited, 0.0015% v/v formaldehyde-treated and 0.5% v/v acetic acid-treated Dunaliella viridis with that of active growing cultures, employing 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenil)-2H-tetrazolium (MTS), another tetrazolium salt.…”

In vitro antiviral activity of antimicrobial peptides against herpes simplex virus 1, adenovirus, and rotavirus

“…Cytotoxicity evaluation -The cell viability was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method (Mossmann 1983, Sieuwerts et al 1995, with minor modifications. Vero, HEp-2, and MA104 cells were grown in 96-well plates for 24 h at 37ºC in a humidified 5% CO 2 atmosphere.…”

Activity of tyrosine kinase inhibitor Dasatinib in neuroblastoma cells in vitro and in orthotopic mouse model

“…After 18 hr, fresh medium or medium containing Dasatinib (10, 100 and 1,000 nM) was added. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay 21 after 72 hr. Expression of beta-galactosidase activity as a marker of cellular senescence was carried out as described.…”