Simultaneous voltage and calcium mapping of genetically purified human induced pluripotent stem cell-derived cardiac myocyte monolayers.
Simultaneous voltage and calcium mapping of genetically purified human induced pluripotent stem cell-derived cardiac myocyte monolayers.
复制标题
基因纯化的人诱导多能干细胞衍生的心肌细胞单层的同步电压和钙图谱。
DOI：
10.1161/circresaha.111.262535
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发表时间：
2012-06-08
影响因子：
20.1
通讯作者：
Herron TJ
中科院分区：
文献类型：
--
作者：
Lee P;Klos M;Bollensdorff C;Hou L;Ewart P;Kamp TJ;Zhang J;Bizy A;Guerrero-Serna G;Kohl P;Jalife J;Herron TJ
人诱导多能干细胞来源的心肌细胞(IPSC-CMS)为研究疾病机制和进行患者特异性药物筛选提供了强大的体外工具。到目前为止，IPSC-CMS的电生理学分析仅限于单细胞记录或小心肌细胞聚集体的低分辨率微电极阵列标测。需要一种新的方法来产生和光学映射大的人IPSC-CM心脏单层的脉冲传播。我们的第一个目标是开发一个多功能的成像平台，用于心脏制剂，包括人IPSC-CM单分子层的多参数电生理标测。我们的第二个目标是创造用于同时测量动作电位和钙波传播的大型电耦合人IPSC-CM单分子膜。开发了一种基于电子控制发光二极管(LED)照明、多波段滤光片和单摄像头传感器的荧光成像平台，并利用该平台同时监测心脏制剂中的动作电位和细胞内钙波传播。然后产生多个大直径(≥1 cm)电耦合的人心脏单层，它们以与通常在啮齿动物心脏单层观察到的相似的速度传播动作电位和钙波。本文介绍的多参数成像系统提供了一种可扩展的使能技术，可以同时测量心脏单层的动作电位和细胞内钙波的幅度和动力学。大规模生产人IPSC-CMS的出现使得现在有可能产生足够数量的均匀的心脏单层，用于研究心律失常的机制，并提供了优于常用的啮齿动物模型的优点。
Human induced pluripotent stem cell derived cardiomyocytes (iPSC-CMs) offer a powerful in-vitro tool to investigate disease mechanisms and to perform patient-specific drug screening. To date electrophysiological analysis of iPSC-CMs has been limited to single cell recordings or low resolution microelectrode array mapping of small cardiomyocyte aggregates. A new method of generating and optically mapping impulse propagation of large human iPSC-CM cardiac monolayers is needed. Our first aim was to develop an imaging platform with versatility for multi-parameter electrophysiological mapping of cardiac preparations, including human iPSC-CM monolayers. Our second aim was to create large electrically coupled human iPSC-CM monolayers for simultaneous action potential and calcium wave propagation measurements. A fluorescence imaging platform based on electronically-controlled light-emitting-diode (LED) illumination, a multi-band emission filter and single camera sensor was developed and utilized to monitor simultaneously action potential and intracellular calcium wave propagation in cardiac preparations. Multiple large diameter (≥1cm) electrically coupled human cardiac monolayers were then generated that propagated action potentials and calcium waves at velocities similar to those commonly observed in rodent cardiac monolayers. The multi-parametric imaging system presented here offers a scalable enabling technology to measure simultaneously action potential and intracellular calcium wave amplitude and dynamics of cardiac monolayers. The advent of large-scale production of human iPSC-CMs makes it possible now to generate sufficient numbers of uniform cardiac monolayers that can be utilized for the study of arrhythmia mechanisms and offers advantages over commonly used rodent models.