In Vitro Cell-Based MTT and Crystal Violet Assays for Drug Toxicity Screening.

The development of novel drug candidates is a current challenge in pharmacology where therapeutic benefits must exceed side effects. Toxicology testing is therefore a fundamental step in drug discovery research. Herein, we describe the first line of toxicology testing program, consisting in cell-based high-throughput screening assays, which have the advantage of being easy, rapid, cheap, and reproducible while providing quantitative information. We illustrate MTT and Crystal Violet assays, two common colorimetric tests able to assess both cytostatic and cytotoxic effects, respectively, of a drug candidate. MTT assay allows evaluation of cellular metabolic activity, by which cell viability can be inferred; Crystal Violet staining is directly correlated with attached viable cells, thus allowing direct assessment of cell survival and death. Therefore, combination of the two methodologies represents a useful tool for predicting drug sensitivity and efficacy, the first milestones in pre-clinical toxicology workflow. © 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Alessandra Gambacurta, Valentina Tullio, Isabella Savini, Maria Valeria Catani, Valeria Gasperi. In Vitro Cell-Based MTT and Crystal Violet Assays for Drug Toxicity Screening. Methods in molecular biology (Clifton, N.J.). 2025;2834:293-301

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PMID: 39312171

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