Földes Gábor, Mioulane Maxime
Imperial Centre for Experimental and Translational Medicine, National Heart and Lung Institute, Imperial College London, London, UK.
Methods Mol Biol. 2013;1052:29-39. doi: 10.1007/7651_2013_25.
Human pluripotent stem cells (hPSC) are investigated as a source of authentic human cardiac cells for drug discovery and toxicological tests. Cell-based assays using automated fluorescence imaging platform and high-content analysis characterize hypertrophic and toxicity profiles of compounds in hPSC-derived cardiomyocytes (hPSC-CM) at the cellular and subcellular levels. In purified population of hPSC-CM loaded with cell tracer probe and cell death markers, both hypertrophic and toxicity profiles can be assessed in live cardiomyocyte cultures. Alternatively, in non-purified cultures of hPSC-CM, hypertrophy, proliferation, and cell death assays can be performed specifically in the cardiomyocyte subpopulation using antibodies directed against cardiac proteins and a combination of cell death- and proliferation-specific fluorescent probes.
人类多能干细胞(hPSC)作为用于药物发现和毒理学测试的真实人类心脏细胞来源而受到研究。使用自动荧光成像平台和高内涵分析的基于细胞的检测方法,在细胞和亚细胞水平上表征了化合物在hPSC衍生的心肌细胞(hPSC-CM)中的肥大和毒性特征。在加载了细胞示踪探针和细胞死亡标记物的hPSC-CM纯化群体中,可以在活心肌细胞培养物中评估肥大和毒性特征。或者,在hPSC-CM的非纯化培养物中,可以使用针对心脏蛋白的抗体以及细胞死亡和增殖特异性荧光探针的组合,在心肌细胞亚群中特异性地进行肥大、增殖和细胞死亡检测。