Cytotoxicity assessment based on the AUC50 using multi-concentration time-dependent cellular response curves
Cytotoxicity assessment based on the AUC50 using multi-concentration time-dependent cellular response curves
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DOI：
10.1016/j.aca.2012.12.047
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发表时间：
2013-02-18
影响因子：
6.2
通讯作者：
Devendran, Vignesh
中科院分区：
文献类型：
--
作者：
Pan, Tianhong;Huang, Biao;Devendran, Vignesh
虽然已经开发了许多指标来量化化学毒性，但大多数指标都存在着实质性的缺陷，如观察时间依赖性强，稳健性不足，无法与阴性对照进行比较。为了评估受试物质的暴露程度，开发了一种名为AUC(50)的细胞毒性测试来描述时间和浓度依赖的细胞反应。通过xCELLigence实时细胞分析高通量(RTCA-HT)系统监测活细胞的动态细胞毒性反应谱，记录并分析细胞数量的变化(命名细胞指数，CI)。引入归一化细胞指数(NCI)以减少实验间差异的影响。考虑了细胞生长的对数阶段，减轻了细胞的自发效应。利用AUC(50)的概念，计算了被测物质的控制线和评估的时间依赖细胞反应曲线(TCRC)之间的面积，并建立了相应的指数杀伤模型(浓度-反应曲线)。通过将HepG2细胞株暴露于七种化合物中，验证了所提出的方法的有效性。我们的研究结果表明，基于AUC的毒性试验可以替代传统的单时间点试验，并有可能成为评估基于细胞的体外试验的常规设置。此外，AUC(50)结合RTCA-HT检测可以实现常规细胞检测无法实现的高通量筛选。(C)2013爱思唯尔B.V.保留所有权利。
Although many indices have been developed to quantify chemical toxicity, substantial shortcoming is inherent in most of them, such as observation time dependence, insufficient robustness, and no comparison with the negative control. To assess the extent of exposure of the tested substance, a cytotoxicity assay named AUC(50) was developed to describe the time and concentration-dependent cellular responses. By monitoring the dynamic cytotoxicity response profile of living cells via the xCELLigence real-time cell analysis high-throughput (RTCA HT) system, changes in cell number (named cell index, CI) were recorded and analyzed subsequently. A normalized cell index (NCI) is introduced to reduce the influence of inter-experimental variations. The log-phase of cellular growth is considered, which alleviates the cell's spontaneous effect. The area between the control line and the assessed time-dependent cellular response curve (TCRC) of the tested substance was calculated, and the corresponding exponential kill model (concentration-response curve) was developed along with exploiting the concept of AUC(50). The validation of the proposed method is demonstrated by exposing HepG2 cell line to seven chemical compounds. Our findings suggested that the proposed AUC-based toxicity assay could be an alternative to the traditional single time-point assay, and it has potential to become routine settings for evaluating the cell-based in vitro assay. Furthermore, the AUC(50) combined with RTCA HT assay can be used to achieve a high-throughput screening that conventional cellular assay cannot achieve. (C) 2013 Elsevier B.V. All rights reserved.