MTT assay to evaluate the cytotoxic potential of a drug




Cell viability, Cytotoxicity, Formazan, MTT assay


Quantification of cell viability and proliferation form the fundamental for numerous in vitro assays in response to external factors. An MTT assay is a colorimetric assay based on assessing the cell metabolic activity. A549 Lung adenocarcinoma cell line was used to see the cytotoxic potential of a new drug for initial screening of apoptosis or necrosis. The biochemical mechanism behind the MTT assay involves NAD(P)H-dependent cellular oxidoreductase enzyme that converts the yellow tetrazolium MTT [3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] into insoluble (E,Z)-5-(4,5-dimethylthiazol-2-yl)-1,3-diphenylformazan (formazan). The formed formazan can be dissolved with dimethyl sulfoxide (DMSO) to give a purple color with characteristic absorption at 540 nm. Intensity of purple color is directly proportional to the cell number and thus indicating the cell viability.

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Author Biographies

Ashutosh Bahuguna, Department of Biotechnology, College of Engineering, Daegu University, Daegu


Mob: +82-10-48321341

Imran Khan, Department of Biotechnology, College of Engineering, Daegu University, Daegu

PhD scholar

Mobile: +821073549718

Vivek K. Bajpai, Department of Applied Microbiology and Biotechnology, Yeungnam University

Assistant Professor

Sun Chul Kang, Department of Biotechnology, College of Engineering, Daegu University, Daegu



Alley MC, Scudiere DA, Monks A, Czerwinski M, Shoemaker R, Boyd MR. Validation of an automated microculture tetrazolium assay (MTA) to assess growth and drug sensitivity of human tumor cell lines. Proc Am Assoc Cancer Res. 1986; 27: 389-91.

Berridge MV, Herst PM, Tan AS. Tetrazolium dyes as tools in cell biology: New insights into their cellular reduction. Biotechnol Annu Rev. 2005; 11: 127-52.

Fotakis G, Timbrell JA. In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett. 2006; 160: 171-77.

Morgan DML. Tetrazolium (MTT) assay for cellular viability and activity. Methods Mol Biol. 1998; 79: 179-84.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65: 5563.

Strober W. Trypan blue exclusion test of cell viability. Curr Protoc Immunol. 2001: 21: 3B:A.3B.1A.3B.2.

Van de Loosdrecht AA, Beelen RH, Ossenkoppele GJ, Broekhoven MG, Langenhuijsen MM. A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. J Immunol Methods. 1994; 174: 311-20.

van Meerloo J, Kaspers GJL, Cloos J. Cell sensitivity assays: The MTT assay. Methods Mol Biol. 2011; 731: 237-45.




How to Cite

Bahuguna, A., I. Khan, V. K. Bajpai, and S. C. Kang. “MTT Assay to Evaluate the Cytotoxic Potential of a Drug”. Bangladesh Journal of Pharmacology, vol. 12, no. 2, Apr. 2017, p. Available Online: Apr 8, 2017, doi:10.3329/bjp.v12i2.30892.



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