Multiparametric High-Content Assays to Measure Cell Health and Oxidative Damage as a Model for Drug-Induced Liver Injury.
Multiparametric High-Content Assays to Measure Cell Health and Oxidative Damage as a Model for Drug-Induced Liver Injury.
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DOI：
10.1002/cpch.90
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发表时间：
2020-12
影响因子：
--
通讯作者：
Markossian S
中科院分区：
文献类型：
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作者：
Pohan G;Espinosa JA;Chen S;Ang KK;Arkin MR;Markossian S
药物性肝损伤是药物研发未获批准以及已批准药物退出市场的重要原因。筛选人肝细胞系的毒性已被广泛用于预测临床前药物开发中药物引起的肝损伤。使用更多样化的标记物评估肝细胞健康状况将增加体外测定的价值，并有助于预测毒性机制。我们描述了三种基于活细胞的检测，使用 HepG2 细胞来测量指示肝毒性的细胞健康参数。第一个测定使用荧光素酶测量细胞 ATP 水平。第二个和第三个测定是多参数高内涵筛选，涵盖一组细胞健康标志物，包括细胞计数、线粒体膜电位和结构、核形态、液泡密度以及活性氧和谷胱甘肽水平。
Drug-induced liver injury is an important cause of non-approval in drug development and the withdrawal of already approved drugs from the market. Screening human hepatic cell lines for toxicity has been used extensively to predict drug-induced liver injury in preclinical drug development. Assessing hepatic-cell health with more diverse markers will increase the value of in vitro assays and help predict the mechanism of toxicity. We describe three live cell-based assays using HepG2 cells to measure cell health parameters indicative of hepatotoxicity. The first assay measures cellular ATP levels using luciferase. The second and third assays are multiparametric high-content screens covering a panel of cell health markers including cell count, mitochondrial membrane potential and structure, nuclear morphology, vacuolar density, and reactive oxygen species and glutathione levels.