Monitoring Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Genetically Encoded Calcium and Voltage Fluorescent Reporters
Monitoring Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes with Genetically Encoded Calcium and Voltage Fluorescent Reporters
复制标题
DOI：
10.1016/j.stemcr.2015.08.009
复制
发表时间：
2015-10-13
影响因子：
5.9
通讯作者：
Gepstein, Lior
中科院分区：
文献类型：
--
作者：
Shinnawi, Rami;Huber, Irit;Gepstein, Lior
人类诱导多能干细胞（hiPSC）技术的出现改变了生物医学研究，为人类疾病建模，药物开发和再生医学提供了新的工具。为了实现其在心血管领域的独特潜力，应该开发有效的方法来对hiPSC衍生的心肌细胞（hiPSC-CM）进行高分辨率、大规模、长期和连续的功能细胞表型分析。为了实现这一目标，我们将hiPSC技术与遗传编码电压（ArcLight）和钙（GCaMP 5G）荧光指示剂相结合。hiPSC-CM中ArcLight和GCaMP 5G的表达允许分别可靠地跟踪跨膜电位和细胞内钙水平的变化。这允许监测动作电位和钙处理特性的短期和长期变化以及对几种药物和来自不同遗传性致心律失常综合征患者的hiPSC-CM的心律失常的发生。将基因编码的荧光报告基因与hiPSC-CM结合可能会为遗传性疾病、发育生物学以及药物开发和测试的研究带来独特的价值。
The advent of the human-induced pluripotent stem cell (hiPSC) technology has transformed biomedical research, providing new tools for human disease modeling, drug development, and regenerative medicine. To fulfill its unique potential in the cardiovascular field, efficient methods should be developed for high-resolution, large-scale, long-term, and serial functional cellular phenotyping of hiPSC-derived cardiomyocytes (hiPSC-CMs). To achieve this goal, we combined the hiPSC technology with genetically encoded voltage (ArcLight) and calcium (GCaMP5G) fluorescent indicators. Expression of ArcLight and GCaMP5G in hiPSC-CMs permitted to reliably follow changes in transmembrane potential and intracellular calcium levels, respectively. This allowed monitoring short- and long-term changes in action-potential and calcium-handling properties and the development of arrhythmias in response to several pharmaceutical agents and in hiPSC-CMs derived from patients with different inherited arrhythmogenic syndromes. Combining genetically encoded fluorescent reporters with hiPSC-CMs may bring a unique value to the study of inherited disorders, developmental biology, and drug development and testing.