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.

Gambacurta, A., Tullio, V., Savini, I., Catani, M.v., Gasperi, V. (2025). In Vitro Cell-Based MTT and Crystal Violet Assays for Drug Toxicity Screening. In Nicolotti O. (a cura di), Computational Toxicology. Nicolotti O. [10.1007/978-1-0716-4003-6_14].

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

Gambacurta, Alessandra;Tullio, Valentina;Savini, Isabella;Catani, Maria Valeria;Gasperi, Valeria
2025-01-01

Abstract

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
Settore BIO/10
Settore BIOS-07/A - Biochimica
English
Rilevanza internazionale
Capitolo o saggio
Absorbance
Colorimetric detection
Crystal Violet
Drug toxicity
IC50 value
In vitro cell-based high-throughput screening assays
MTT
Gambacurta, A., Tullio, V., Savini, I., Catani, M.v., Gasperi, V. (2025). In Vitro Cell-Based MTT and Crystal Violet Assays for Drug Toxicity Screening. In Nicolotti O. (a cura di), Computational Toxicology. Nicolotti O. [10.1007/978-1-0716-4003-6_14].
Gambacurta, A; Tullio, V; Savini, I; Catani, Mv; Gasperi, V
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/391965
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