Goedel Alexander, Zawada Dorota M, Zhang Fangfang, Chen Zhifen, Moretti Alessandra, Sinnecker Daniel
Medical Department I, University Hospital Klinikum rechts der Isar, Technical University of Munich; German Centre for Cardiovascular Research (DZHK), Munich Heart Alliance.
Medical Department I, University Hospital Klinikum rechts der Isar, Technical University of Munich.
J Vis Exp. 2018 Sep 27(139):58134. doi: 10.3791/58134.
Cardiomyocytes generated from human induced pluripotent stem cells (iPSC-CMs) are an emerging tool in cardiovascular research. Rather than being a homogenous population of cells, the iPSC-CMs generated by current differentiation protocols represent a mixture of cells with ventricular-, atrial-, and nodal-like phenotypes, which complicates phenotypic analyses. Here, a method to optically record action potentials specifically from ventricular-like iPSC-CMs is presented. This is achieved by lentiviral transduction with a construct in which a genetically-encoded voltage indicator is under the control of a ventricular-specific promoter element. When iPSC-CMs are transduced with this construct, the voltage sensor is expressed exclusively in ventricular-like cells, enabling subtype-specific optical membrane potential recordings using time-lapse fluorescence microscopy.
源自人诱导多能干细胞的心肌细胞（iPSC-CMs）是心血管研究中一种新兴的工具。通过当前分化方案产生的iPSC-CMs并非同质细胞群体，而是代表具有心室样、心房样和结样表型的细胞混合物，这使得表型分析变得复杂。在此，介绍一种专门从心室样iPSC-CMs光学记录动作电位的方法。这是通过慢病毒转导一种构建体来实现的，在该构建体中，一种基因编码的电压指示剂受心室特异性启动子元件的控制。当用这种构建体转导iPSC-CMs时，电压传感器仅在心室样细胞中表达，从而能够使用延时荧光显微镜进行亚型特异性的光学膜电位记录。
