Abstract
In the course of examining the effects on cells of polyamines, their metabolites, and polyamine analogs, it is often necessary to make some measure of cellular activity as an indicator of cell damage or cytotoxicity. One of the simplest assays utilizes 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), a water-soluble yellow dye that is readily taken up by viable cells and reduced by the action of mitochondrial dehydrogenases (1). The reduction product is a water-insoluble blue formazan, that must then be dissolved for calorimetric measurement. Ethanol or propanol (2), acid-isopropanol (0.04M HCl in propan-2-ol) (3), acid-isopropanol plus 10% Triton X-100 (4), mineral oil (unspecified), or dimethyl sulfoxide (DMSO) (5) have all been suggested. In the author’s hands DMSO was found to be the most satisfactory (Fig. 1).
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References
Slater, T. F., Sawyer, B., and Strauli, U. (1963) Studies on succinate-tetrazolium reductase systems III Points of coupling of four different tetrazolium salts. Biochim. Biophys. Acta 77, 383–393.
Denizot, F. and Lang, R. (1986) Rapid colorimetric assay for cell growth and survival: modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immunol. Methods 89, 271–277.
Mossman, T. (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65, 55–63.
Takeuchi, H., Baba, M., and Shigeta, S. (1991) An application of tetrazolium (MTT) colorimetric assay for the screening of anti-herpes simplex virus compounds. J. Virol. Methods 33, 61–71.
Carmichael, J., DeGraff, W. G., Gazdar, A. F., Minna, J. D., and Mitchell, J. B. (1987) Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47, 936–942.
Vestica, D. T., Skehan, P., Scudiero, D., Monks, A., Pittman, A., and Boyd, M. R. (1991) Tetrazolium-based assays for cellular viability, a critical examination of selected parameters affecting formazan production. Cancer Res. 51, 2515–2520.
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© 1998 Humana Press Inc.
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Morgan, D.M.L. (1998). Tetrazolium (MTT) Assay for Cellular Viability and Activity. In: Morgan, D.M.L. (eds) Polyamine Protocols. Methods in Molecular Biology™, vol 79. Humana Press. https://doi.org/10.1385/0-89603-448-8:179
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DOI: https://doi.org/10.1385/0-89603-448-8:179
Publisher Name: Humana Press
Print ISBN: 978-0-89603-448-8
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