High throughput, non-invasive and dynamic toxicity screening on adherent cells using respiratory measurements
High throughput, non-invasive and dynamic toxicity screening on adherent cells using respiratory measurements
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DOI：
10.1016/j.tiv.2009.04.018
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发表时间：
2010-03-01
影响因子：
3.2
通讯作者：
Heinzle, Elmar
中科院分区：
文献类型：
--
作者：
Beckers, Simone;Noor, Fozia;Heinzle, Elmar
提出了一种基于氧的发光衰减时间检测的高通量毒理学评价的动态呼吸分析方法。该方法使用24孔板（OxoDish），在置于加湿CO2培养箱中的传感器皿读数器的帮助下进行读数。贴壁原代大鼠肝细胞和人肝细胞系Hep G2暴露于已知的毒性化合物。溶解氧浓度是呼吸的量度，用固定在每个孔中心的氧传感器光电二极管测量。在测定期间，将细胞保持在培养皿中，并且之后可以进行处理以用于进一步分析。这种动态、非侵入性测量允许计算任何孵育时间点的50%致死浓度（LC 50），其给出浓度-时间依赖性响应，而无需进一步操作或从孵育器中取出细胞。将毒代动力学特征与磺酰罗丹明B试验（一种常见的细胞毒性试验）进行比较。新的测定法是稳健的和灵活的，非常容易进行，并提供反映动态毒性反应的连续在线呼吸数据。它可以适用于任何基于细胞的系统，计算的动力学有助于理解细胞死亡机制。(C)2009爱思唯尔有限公司保留所有权利。
A dynamic respiration assay based on luminescence decay time detection of oxygen for high throughput toxicological assessment is presented. The method uses 24-well plates (OxoDishes) read with the help of a sensor dish reader placed in a humidified CO2-incubator. Adherent primary rat hepatocytes and the human hepatic cell line Hep G2 were exposed to known toxic compounds. Dissolved oxygen concentration, a measure of respiration, was measured with an oxygen sensor optode immobilized in the centre of each well. The cells were maintained in the dishes during the assay period and can afterwards be processed for further analyses. This dynamic, non-invasive measurement allowed calculation of 50% lethal concentrations (LC50) for any incubation time point giving concentration-time-dependent responses without further manipulation or removal of the cells from the incubator. Toxicokinetic profiles are compared with Sulforhodamine B assay, a common cytotoxicity assay. The novel assay is robust and flexible, very easy to carry out and provides continuous online respiration data reflecting dynamic toxicity responses. It can be adapted to any cell-based system and the calculated kinetics contributes to understanding of cell death mechanisms. (C) 2009 Elsevier Ltd. All rights reserved.