The rainbow trout gill cell line (RTgill-W1), via test guideline 249 of the Organisation for Economic Co-operation and Development, has been established as a promising New Approach Methodology, although to advance confidence in the method more case studies are needed that: 1) expand our understanding of applicability domains (chemicals with diverse properties); 2) increase methodological throughput (96-well format); and 3) demonstrate biological relevance (in vitro to in vivo comparisons; gill vs. other cells). Accordingly, the objective of our study was to characterize the cytotoxicity of 19 pesticides against RTgill-W1 cells, and also liver (RTL-W1) and gut epithelial (RTgutGC) cell lines, and then to compare the in vitro and in vivo data. Of the 19 pesticides tested, 11, 9, and 8 were cytotoxic to the RTgill-W1, RTL-W1, and RTgutGC cells, respectively. Six pesticides (carbaryl, chlorothalonil, chlorpyrifos, dimethenamid-P, metolachlor, and S-metolachlor) were cytotoxic to all three cell lines. Aminomethylphosphonic acid, chlorantraniliprole, dicamba, diquat, imazethapyr, and permethrin exhibited cell-line–specific toxicity. No cytotoxic responses were observed for three herbicides (atrazine, glyphosate, and metribuzin) and four insecticides (clothianidin, diazinon, imidacloprid, and thiamethoxam). When cytotoxicity was measured, there was a strong correlation (r_s = 0.9, p < 0.0001) between in vitro median effect concentration (EC50) values (based on predicted concentrations using the In Vitro Mass Balance Model Equilibrium Partitioning (IV-MBM EQP) Ver. 2.1) derived from RTgill-W1 and RTL-W1 cells with in vivo median lethal concentration (LC50) values from 96-h acute toxicity studies with trout. In all 28 cases, the in vitro EC50 was within 18-fold of the in vivo LC50. These data help increase our understanding of the ecotoxicological domains of applicability for in vitro studies using cultured rainbow trout cells, while also demonstrating that these assays performed well in a 96-well format and have promise to yield data of biological relevance. Environ Toxicol Chem 2024;00:1–13. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

中文翻译：

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19 种农药对虹鳟鱼鳃、肝和肠细胞系的细胞毒性

虹鳟鱼鳃细胞系 (RTgill-W1)，通过经济合作与发展组织的测试指南 249，已被确立为一种有前途的新方法方法，尽管为了提高对该方法的信心，需要更多的案例研究：1）扩大我们对适用领域（具有不同特性的化学品）的理解； 2) 提高方法学通量（96 孔格式）； 3) 证明生物学相关性（体外与体内比较；鳃细胞与其他细胞）。因此，我们研究的目的是表征 19 种农药对 RTgill-W1 细胞以及肝 (RTL-W1) 和肠上皮 (RTgutGC) 细胞系的细胞毒性，然后比较体外和体内数据。在测试的 19 种农药中，分别有 11 种、9 种和 8 种对 RTgill-W1、RTL-W1 和 RTgutGC 细胞具有细胞毒性。六种农药（西维因、百菌清、毒死蜱、噻吩草胺、异丙甲草胺和精异丙甲草胺）对所有三种细胞系均具有细胞毒性。氨甲基膦酸、氯虫苯甲酰胺、麦草畏、敌草快、咪草烟和氯菊酯表现出细胞系特异性毒性。三种除草剂（莠去津、草甘膦和赛克津）和四种杀虫剂（噻虫胺、二嗪农、吡虫啉和噻虫嗪）未观察到细胞毒性反应。测量细胞毒性时，_体外中位效应浓度 (EC50) 值之间存在很强的相关性（rs  = 0.9，p  < 0.0001）（基于使用体外质量平衡模型平衡分配 (IV-MBM EQP) 预测的浓度）版本 2.1）源自 RTgill-W1 和 RTL-W1 细胞，体内半数致死浓度 (LC50) 值来自鳟鱼的 96 小时急性毒性研究。在所有 28 例中，体外 EC50 均在体内 LC50 的 18 倍以内。这些数据有助于加深我们对使用培养虹鳟细胞进行体外研究的生态毒理学领域的了解，同时还证明这些测定在 96 孔格式中表现良好，并有望产生生物学相关的数据。环境毒理学2024；00：1–13。 © 2023 作者。《环境毒理学和化学》由 Wiley periodicals LLC 代表 SETAC 出版。