Frankel Cardiovascular Regeneration Core Laboratory, Department of Internal Medicine, Division of Cardiovascular Medicine.
Center for Arrhythmia Research.
JCI Insight. 2021 May 24;6(10):134368. doi: 10.1172/jci.insight.134368.
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have been used extensively to model inherited heart diseases, but hiPSC-CM models of ischemic heart disease are lacking. Here, our objective was to generate an hiPSC-CM model of ischemic heart disease. To this end, hiPSCs were differentiated into functional hiPSC-CMs and then purified using either a simulated ischemia media or by using magnetic antibody-based purification targeting the nonmyocyte population for depletion from the cell population. Flow cytometry analysis confirmed that each purification approach generated hiPSC-CM cultures that had more than 94% cTnT+ cells. After purification, hiPSC-CMs were replated as confluent syncytial monolayers for electrophysiological phenotype analysis and protein expression by Western blotting. The phenotype of metabolic stress-selected hiPSC-CM monolayers recapitulated many of the functional and structural hallmarks of ischemic CMs, including elevated diastolic calcium, diminished calcium transient amplitude, prolonged action potential duration, depolarized resting membrane potential, hypersensitivity to chemotherapy-induced cardiotoxicity, depolarized mitochondrial membrane potential, depressed SERCA2a expression, reduced maximal oxygen consumption rate, and abnormal response to β1-adrenergic receptor stimulation. These findings indicate that metabolic selection of hiPSC-CMs generated cell populations with phenotype similar to what is well known to occur in the setting of ischemic heart failure and thus provide a opportunity for study of human ischemic heart disease.
人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）已被广泛用于模拟遗传性心脏病，但缺血性心脏病的 hiPSC-CM 模型仍然缺乏。在这里，我们的目标是生成缺血性心脏病的 hiPSC-CM 模型。为此，将 hiPSC 分化为功能性 hiPSC-CM，然后使用模拟缺血介质或使用针对非心肌细胞群的基于磁性抗体的纯化方法从细胞群中去除来进行纯化。流式细胞术分析证实，每种纯化方法都产生了超过 94% cTnT+细胞的 hiPSC-CM 培养物。纯化后，将 hiPSC-CM 重新铺板为融合的合胞单层，以进行电生理表型分析和通过 Western blot 进行蛋白质表达分析。代谢应激选择的 hiPSC-CM 单层的表型再现了许多缺血性 CM 的功能和结构特征，包括升高的舒张钙、钙瞬变幅度降低、动作电位持续时间延长、静息膜电位去极化、对化疗诱导的心脏毒性敏感性增加、线粒体膜电位去极化、SERCA2a 表达降低、最大耗氧量降低以及对β1-肾上腺素能受体刺激的异常反应。这些发现表明，代谢选择 hiPSC-CM 生成的细胞群体具有与已知在缺血性心力衰竭情况下发生的表型相似的表型，从而为研究人类缺血性心脏病提供了机会。
