General Cytotoxicity Assessment by Means of the MTT Assay
Cytotoxicity assays were among the first in vitro bioassay methods used to predict toxicity of substances to various tissues. In vitro cytotoxicity testing provides a crucial means for safety assessment and screening, and for ranking compounds. The choice of using a particular cytotoxicity assay technology may be influenced by specific research goals. As such, four main classes of assays are used to monitor the response of cultured cells after treatment with potential toxicants. These methods measure viability, cell membrane integrity, cell proliferation, and metabolic activity. In this chapter, we focus on the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium reduction colorimetric assay to evaluate detrimental intracellular effects on metabolic activity. This assay is well-characterized, simple to use and remains popular in several laboratories worldwide.
Key wordsCytotoxicity Tetrazolium salt IC50 Balb/c 3T3 cells Rat hepatocytes
The authors acknowledge financial support from the ALIVE Foundation. Laia Tolosa was a recipient of a Sara Borrell Contract from the “Instituto de Salud Carlos III” of the Spanish Ministry of Economy and Competitiveness.
- 1.Gomez-Lechon MJ, Ponsoda X, Castell JV (2000) In vitro toxicity testing. In: Doyle A, Griffiths JB (eds) Cell and tissue culture for medical research. Wiley, Chinchester, pp 402–419Google Scholar
- 5.Freshney R (1987) Culture of animal cells: a manual of basic technique. Alan R. Liss, Inc., New YorkGoogle Scholar
- 22.Ponsoda X, Gomez-Lechon MJ, Castell JV (1998) Toxicity and cell density monitoring in monolayer and three-dimmensional cultures with the XTT assay. ATLA 26:331–342Google Scholar
- 43.Tan AS, Berridge MV (2000) Superoxide produced by activated neutrophils efficiently reduces the tetrazolium salt, WST-1 to produce a soluble formazan: a simple colorimetric assay for measuring respiratory burst activation and for screening anti-inflammatory agents. J Immunol Methods 238:59–68PubMedCrossRefGoogle Scholar