Abstract
The MTT (3-[4,5-dimethylthiazol-2yl]-2,5-diphenyl tetrazolium bromide) growth assay developed by Mosmann (1) offers a simple, rapid, and precise measurement of cell viability and proliferation of adherent cell lines (2). The value of this assay is in the screening of large numbers of samples. The MTT assay, a quantitative colorimetric assay is based on the living cell’s ability to reduce the tetrazolium salt MTT, a pale yellow substrate to a dark-blue formazan product. The mitochondrial succinate-dehydrogenases (3) of viable cells cleave the tetrazolium ring in active mitochondria into formazan crystals. The crystals can be dissolved in acid isopropyl alcohol, mineral oil (4), or dimethyl sulfoxide (DMSO) (5). The resulting blue solution can be measured semiautomatically using a scanning multiwell spectrophotometer. Our laboratory has successfully applied the MTT-based growth assay with some modifications (6) to investigate the growth effects of human cytokines on HOC cell lines (7), but we have to keep its limitations and pitfalls in mind (see Note 1). For use in tests of floating cell lines, the MTT assay may be less optimal. Using this assay for screening of primary tumor samples may produce limited results, because cell contaminants may result in high-background values (4). However, accepting the limitations of the MTT assay, the optimum assay conditions have to be selected and adapted to the cell lines that are under investigation. The MTT-based growth assay, as described in this chapter, is a reliable and sensitive test for the determination of cell growth of human ovarian-carcinoma cells.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Mosmann, T. (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Meth. 65, 55–63.
Scudiero, D. A., Shoemaker, R. H., Paull, K. D., Monks, A., Tierney, S., Nofziger, T. H., et al. (1988) Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other cell lines. Cancer Res. 48, 4827–4833.
Slater, T. F., Sawyer, B., and Strauli, U. (1963) Studies on succinate-tetrazolium reductase system III. Points of coupling of four different tetrazolium salts. Biochem. Biophys. Acta 77, 383–393.
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.
Park, J-G., Kramer, B. S., Steinberg, S. M., Carmichael, J., Collins, J. M., Minna, J. D., et al. (1987) Chemosensitivity testing of human colorectal carcinoma cell lines using a tetrazolium-based colorimetric assay. Cancer Res. 447, 5875–5879.
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. Meth. 89, 271–277.
Spinner, D. M., Brandstetter, T., Kiechle-Schwarz, M., du Bois, A., Angel, P., and Bauknecht, T. (1995) C-jun expression and growth stimulation in human ovarian carcinoma cell lines following exposure to cytokines. Int. J. Cancer 63, 423–427.
Vistica, 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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Spinner, D.M. (2000). MTT Growth Assays in Ovarian Cancer. In: Bartlett, J.M.S. (eds) Ovarian Cancer. Methods in Molecular Medicine™, vol 39. Humana Press. https://doi.org/10.1385/1-59259-071-3:175
Download citation
DOI: https://doi.org/10.1385/1-59259-071-3:175
Publisher Name: Humana Press
Print ISBN: 978-0-89603-583-6
Online ISBN: 978-1-59259-071-1
eBook Packages: Springer Protocols