Validation of cell impedance sensing as a high-throughput approach for toxicity evaluation in vitro

作为毒理学领域的世界领导者，查尔斯河实验室（CRL）正在寻求通过开发新的基于细胞的检测来扩大其毒理学专业知识。在本项目中，我们提出应用电细胞阻抗传感技术（ECIS）来评估与稳态的两个关键参数相关的功能终点，即内皮细胞通透性和血小板聚集，内皮细胞和血小板在稳态和血栓形成中起着关键作用，并且药物的血管安全性已经成为药物开发和药物批准过程中的重要标准。通过测量器官中的伊文思蓝内皮外渗，在动物模型中常规测试药物诱导的内皮渗漏。血小板功能可以使用光透射聚集测定法或阻抗聚集测定法离体评价，阻抗聚集测定法测量血小板在一对浸入全血样品中的电极上的粘附。虽然非常简单和敏感，这两个标准化的体内程序是非常低的吞吐量的性质，这使得他们迅速不切实际和昂贵的，如果一个aimsat建立剂量-毒性曲线早在选择有前途的铅候选化合物库。为了解决这一局限性，我们将验证两种基于细胞的检测方法，使用96孔板的电阻抗技术来定量药物诱导的内皮细胞通透性和血小板聚集。

As a world leader in the field of toxicology, Charles River Laboratories (CRL) is seeking to expand itsportfolio of toxicology expertises through the development of novel cell-based assays. In this project, wepropose to apply electric cell impedance sensing (ECIS) for the evaluation of functional end-point associatedwith two critical parameters of homeostasis, that is endothelium permeability and platelet aggregation.Endothelial cells and platelets play pivotal roles in homeostasis and thrombosis and it has become clear thatvascular safety of drugs constitutes an important criterion in both the drug development and the drug approvalprocesses. Drug-induced endothelium leakage is routinely tested in animal models by measuring Evans Blueendothelium extravasation in organs. Platelet function can be evaluated ex vivo using light transmissionaggregation assay or by impedance aggregometry which measure platelet adhesion on a pair of electrodeimmersed in a whole blood sample. Although very simple and sensitive, theses two standardised in vivoprocedures are very low throughput in nature, which makes them rapidly unpractical and expensive if one aimsat establishing dose-toxicity curves early in the selection of promising lead candidates from compoundlibraries. To address this limitation, we will validate two cell-based assays using an electrical impedancetechnology in 96-well format to quantify drugs-induced endothelium permeability and platelet aggregation.