Antioxidant compounds interfere with the 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide cytotoxicity assay

M. Natarajan, S. Mohan, B. R. Martinez, M. L. Meltz, T. S. Herman

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

Antioxidants are often added to culture media as cytoprotective agents. We examined the effects of antioxidants on the results and interpretation of the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay for cell viability. Without cells, the thiol-containing antioxidant compounds β-mercaptoethanol, dithiothreitol, pyrrolidine-dithiocarbamate, and N-acetyl-L-cysteine (acetylcysteine) reduced MTT tetrazolium salts to a blue formazan product in a dose-dependent manner. Addition of the compounds L-ascorbic acid and (+)α-tocopherol acid succinate had different effects. In contrast, addition of the antioxidants N-acetyl-5methoxytryptamine and (-)-2-oxo-4-thiazolidine carboxylic acid, which do not contain reactive thiol groups, did not result in the development of blue formazan product. These results showed that antioxidants, and potentially other chemotherapeutic compounds that contain free thiol groups or other reducing equivalents, readily reduce MTT to produce the blue formazan, irrespective of the viability of the cells present. This undescribed reaction can, therefore, significantly influence the results and interpretation of cell-viability experiments.

Original languageEnglish (US)
Pages (from-to)405-414
Number of pages10
JournalCancer Detection and Prevention
Volume24
Issue number5
StatePublished - Dec 27 2000

Keywords

  • Antioxidants
  • Cytotoxicity
  • Drugs
  • In vitro
  • MTT assay

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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