Zhang Hong-Yuan, Zhang Ting-Ting, Machuki Jeremiah Ong'achwa, Wu Li-Juan, Sun Hong
Department of Physiology, Xuzhou Medical University, Xuzhou 221004, China.
Institute of Cardiovascular Disease Research, Xuzhou Medical University, Xuzhou 221004, China.
Sheng Li Xue Bao. 2018 Jun 25;70(3):281-286.
The present study was aimed to characterize the electrophysiology of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). IMR90-4 cells were induced to differentiate into cardiomyocytes by temporal modulation of regulators of canonical Wnt signaling. The protein expression of cardiac troponin T (cTnT) was detected by immunofluorescence staining and flow cytometry, and the differentiation rate of hiPSC-CMs was calculated. The action potentials (APs) of hiPSC-CMs were recorded by patch clamp and used to classify different types of cardiomyocytes. The electrophysiological characteristics of hiPSC-CMs were further analyzed. The results showed that the cTnT positive rate of hiPSC-CMs was above 95%. hiPSC-CMs were differentiated into 3 types of cardiomyocytes based on the properties of AP: ventricular-, atrial- and nodal-like cells. In comparison with the other two types of cells, the APs of ventricular-like cells exhibited longer duration, higher amplitude and higher dV/dt. The nodal-like cells had the lowest dV/dt among all the three types. These results indicate that hiPSC can be differentiated into the cardiomyocytes with high purity and the differentiated hiPSC-CMs have similar electrophysiological characteristics to adult cardiomyocytes.
本研究旨在表征人诱导多能干细胞衍生心肌细胞（hiPSC-CMs）的电生理学特性。通过对经典Wnt信号通路调节因子进行时间调控，诱导IMR90-4细胞分化为心肌细胞。通过免疫荧光染色和流式细胞术检测心肌肌钙蛋白T（cTnT）的蛋白表达，并计算hiPSC-CMs的分化率。采用膜片钳记录hiPSC-CMs的动作电位（APs），并用于对不同类型的心肌细胞进行分类。进一步分析hiPSC-CMs的电生理特性。结果显示，hiPSC-CMs的cTnT阳性率在95%以上。基于AP的特性，hiPSC-CMs分化为3种类型的心肌细胞：心室样、心房样和结样细胞。与其他两种类型的细胞相比，心室样细胞的APs持续时间更长、幅度更高、dV/dt更高。在所有三种类型中，结样细胞的dV/dt最低。这些结果表明，hiPSC可以分化为高纯度的心肌细胞，且分化后的hiPSC-CMs具有与成年心肌细胞相似的电生理特性。
