This paper proposes a new non-invasive, low-cost, and fully automated platform to quantitatively analyze dynamics of human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) at the single-cell level by holographic image-based tracking for cardiotoxicity screening. A dense Farneback optical flow method and holographic imaging informatics were combined to characterize the contractile motion of a single CM, which obviates the need for costly equipment to monitor a CM's mechanical beat activity. The reliability of the proposed platform was tested by single-cell motion characterization, synchronization analysis, motion speed measurement of fixed CMs versus live CMs, and noise sensitivity. The applicability of the motion characterization method was tested to determine the pharmacological effects of two cardiovascular drugs, isoprenaline (166 nM) and E−4031 (500 μM). The experiments were done using single CMs and multiple cells, and the results were compared to control conditions. Cardiomyocytes responded to isoprenaline by increasing the action potential (AP) speed and shortening the resting period, thus increasing the beat frequency. In the presence of E−4031, the AP speed was decreased, and the resting period was prolonged, thus decreasing the beat frequency. The findings offer insights into single hiPS-CMs’ contractile motion and a deep understanding of their kinetics at the single-cell level for cardiotoxicity screening.

本文提出了一种新的非侵入性、低成本和全自动平台，通过基于全息图像的心脏毒性跟踪在单细胞水平上定量分析人诱导多能干细胞衍生心肌细胞 (hiPS-CMs) 的动态。筛选。密集的 Farneback 光流方法和全息成像信息学相结合来表征单个 CM 的收缩运动，从而无需昂贵的设备来监测 CM 的机械节拍活动。所提出平台的可靠性通过单细胞运动表征、同步分析、固定 CM 与实时 CM 的运动速度测量以及噪声敏感性进行了测试。测试了运动表征方法的适用性，以确定两种心血管药物的药理作用，异丙肾上腺素 (166 nM) 和 E-4031 (500 μM)。使用单个 CM 和多个细胞进行实验，并将结果与​​对照条件进行比较。心肌细胞通过增加动作电位 (AP) 速度和缩短静息期来响应异丙肾上腺素，从而增加搏动频率。在 E-4031 的存在下，AP 速度降低，休息时间延长，从而降低搏动频率。这些发现提供了对单个 hiPS-CM 收缩运动的见解，并深入了解了它们在单细胞水平上用于心脏毒性筛查的动力学。心肌细胞通过增加动作电位 (AP) 速度和缩短静息期来响应异丙肾上腺素，从而增加搏动频率。在 E-4031 的存在下，AP 速度降低，休息时间延长，从而降低搏动频率。这些发现提供了对单个 hiPS-CM 收缩运动的见解，并深入了解了它们在单细胞水平上用于心脏毒性筛查的动力学。心肌细胞通过增加动作电位 (AP) 速度和缩短静息期来响应异丙肾上腺素，从而增加搏动频率。在 E-4031 的存在下，AP 速度降低，休息时间延长，从而降低搏动频率。这些发现提供了对单个 hiPS-CM 收缩运动的见解，并深入了解了它们在单细胞水平上用于心脏毒性筛查的动力学。