Cardiovascular Research Institute, Division of Cardiovascular Diseases, Department of Internal Medicine, Kansas City, USA, KS.
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, USA, KS.
Acta Pharmacol Sin. 2018 Oct;39(10):1590-1603. doi: 10.1038/aps.2017.183. Epub 2018 Apr 5.
Induced pluripotent stem cell (iPSC)-based cardiac regenerative medicine requires the efficient generation, structural soundness and proper functioning of mature cardiomyocytes, derived from the patient's somatic cells. The most important functional property of cardiomyocytes is the ability to contract. Currently available methods routinely used to test and quantify cardiomyocyte function involve techniques that are labor-intensive, invasive, require sophisticated instruments or can adversely affect cell vitality. We recently developed optical flow imaging method analyses and quantified cardiomyocyte contractile kinetics from video microscopic recordings without compromising cell quality. Specifically, our automated particle image velocimetry (PIV) analysis of phase-contrast video images captured at a high frame rate yields statistical measures characterizing the beating frequency, amplitude, average waveform and beat-to-beat variations. Thus, it can be a powerful assessment tool to monitor cardiomyocyte quality and maturity. Here we demonstrate the ability of our analysis to characterize the chronotropic responses of human iPSC-derived cardiomyocytes to a panel of ion channel modulators and also to doxorubicin, a chemotherapy agent with known cardiotoxic side effects. We conclude that the PIV-derived beat patterns can identify the elongation or shortening of specific phases in the contractility cycle, and the obtained chronotropic responses are in accord with known clinical outcomes. Hence, this system can serve as a powerful tool to screen the new and currently available pharmacological compounds for cardiotoxic effects.
基于诱导多能干细胞(iPSC)的心脏再生医学需要从患者体细胞中高效产生、结构健全且功能正常的成熟心肌细胞。心肌细胞最重要的功能特性是收缩能力。目前,常用于测试和量化心肌细胞功能的常用方法涉及到劳动强度大、侵袭性强、需要复杂仪器或可能对细胞活力产生不利影响的技术。我们最近开发了一种光流成像方法,可以在不影响细胞质量的情况下,从视频显微镜记录中分析和量化心肌细胞的收缩动力学。具体来说,我们对高帧率的相差视频图像进行自动粒子图像测速(PIV)分析,得出可用于描述跳动频率、幅度、平均波形和跳动间变化的统计量。因此,它可以成为监测心肌细胞质量和成熟度的有力评估工具。在这里,我们证明了我们的分析能够描述人 iPSC 衍生的心肌细胞对一系列离子通道调节剂以及具有已知心脏毒性副作用的化疗药物阿霉素的变时反应。我们得出结论,PIV 衍生的跳动模式可以识别收缩周期中特定相位的伸长或缩短,并且获得的变时反应与已知的临床结果一致。因此,该系统可作为筛选新的和现有的心脏毒性化合物的有力工具。