Integrated Analysis of Contractile Kinetics, Force Generation, and Electrical Activity in Single Human Stem Cell-Derived Cardiomyocytes.
Integrated Analysis of Contractile Kinetics, Force Generation, and Electrical Activity in Single Human Stem Cell-Derived Cardiomyocytes.
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DOI：
10.1016/j.stemcr.2015.10.017
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发表时间：
2015-12-08
影响因子：
5.9
通讯作者：
Domian IJ
中科院分区：
文献类型：
--
作者：
Kijlstra JD;Hu D;Mittal N;Kausel E;van der Meer P;Garakani A;Domian IJ
心肌细胞功能的定量分析是以干细胞为基础的人体心脏生理学和病理生理学体外研究的基础。我们提出了一种通过同时定量分析收缩动力学、力产生和电活动的方法来综合评价单个人多能干细胞来源的心肌细胞(hPSC-CMS)的功能。我们证明，对收缩hPSC-CMS的电影的统计分析可以用于量化细胞形态随时间的变化，并计算收缩动力学。使用结合底物硬度的生物力学模型，我们计算了单细胞分辨率下的心肌细胞力产生，并用传统的牵引力显微镜验证了这种方法。通过添加荧光钙指示剂或膜电位染料，可以同时分析收缩性能和钙处理或动作电位形态。因此，我们的方法在心脏病研究、药物开发和心脏毒性筛选方面具有广泛的应用潜力。一种无偏量化心肌细胞收缩动力学的方法将生物力学模型用于力产生的分析与钙处理和膜电位的荧光分析完全兼容具有药物筛选潜力的集成机电方法Domian及其同事提出了一种在单细胞分辨率下同时评估干细胞来源的心肌细胞的收缩动力学、力产生和电活动的新方法。该方法识别在柔性底物上收缩的心肌细胞的形态变化，结合生物力学模型来计算力，并与用于药物筛选的钙循环和膜电位的荧光分析相兼容。
The quantitative analysis of cardiomyocyte function is essential for stem cell-based approaches for the in vitro study of human cardiac physiology and pathophysiology. We present a method to comprehensively assess the function of single human pluripotent stem cell-derived cardiomyocyte (hPSC-CMs) through simultaneous quantitative analysis of contraction kinetics, force generation, and electrical activity. We demonstrate that statistical analysis of movies of contracting hPSC-CMs can be used to quantify changes in cellular morphology over time and compute contractile kinetics. Using a biomechanical model that incorporates substrate stiffness, we calculate cardiomyocyte force generation at single-cell resolution and validate this approach with conventional traction force microscopy. The addition of fluorescent calcium indicators or membrane potential dyes allows the simultaneous analysis of contractility and calcium handling or action potential morphology. Accordingly, our approach has the potential for broad application in the study of cardiac disease, drug discovery, and cardiotoxicity screening. An unbiased methodology for quantification of cardiomyocyte contractile kinetics Incorporation of a biomechanical model for the analysis of force generation Fully compatible with fluorescent assays of calcium handling and membrane potential An integrated electromechanical approach with potential for drug screening assays Domian and colleagues present a novel methodology for the concurrent assessment of contractile kinetics, force generation, and electrical activity of stem cell-derived cardiomyocytes at single-cell resolution. The approach identifies morphological changes in cardiomyocytes contracting on flexible substrates, incorporates a biomechanical model to calculate force, and is compatible with fluorescent assays of calcium cycling and membrane potential for drug screening.