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Neutral Red Uptake Assay to Assess Cytotoxicity In Vitro

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Cell Viability Assays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2644))

Abstract

The neutral red uptake (NRU) assay is a cell viability assay that can be used for the assessment of compound-induced cytotoxicity. It is based on the ability of living cells to incorporate neutral red, a weak cationic dye, in lysosomes. The quantification of xenobiotic-induced cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red when compared to cells exposed to corresponding vehicle controls. The NRU assay is mainly used for hazard assessment in in vitro toxicology applications. Hence, this method has been incorporated in regulatory recommendations such as the OECD test guideline TG 432, in which an in vitro 3T3-NRU-phototoxicityassay is described to assess the cytotoxicity of compounds in the presence or absence of UV light.

This book chapter describes a detailed protocol to carry out the NRU assay using the human hepatoma cell line HepG2, which is frequently employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetylsalicylic acid is assessed.

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Correspondence to Robim M. Rodrigues .

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Rodrigues, R.M., Stinckens, M., Ates, G., Vanhaecke, T. (2023). Neutral Red Uptake Assay to Assess Cytotoxicity In Vitro. In: Friedrich, O., Gilbert, D.F. (eds) Cell Viability Assays. Methods in Molecular Biology, vol 2644. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3052-5_15

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  • DOI: https://doi.org/10.1007/978-1-0716-3052-5_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3051-8

  • Online ISBN: 978-1-0716-3052-5

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