Häkli Martta, Kreutzer Joose, Mäki Antti-Juhana, Välimäki Hannu, Cherian Reeja Maria, Kallio Pasi, Aalto-Setälä Katriina, Pekkanen-Mattila Mari
Heart Group, Faculty of Medicine and Health Technology, Tampere University, Tampere 33520, Finland.
Micro- and Nanosystems Research Group, Faculty of Medicine and Health Technology, Tampere University, Tampere 33720, Finland.
Stem Cells Int. 2022 Dec 19;2022:9438281. doi: 10.1155/2022/9438281. eCollection 2022.
Ischemic heart disease is the most common cardiovascular disease and a major burden for healthcare worldwide. However, its pathophysiology is still not fully understood, and human-based models for disease mechanisms and treatments are needed. Here, we used human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to model acute ischemia-reperfusion in our novel cell culture assembly. The assembly enables exchange of oxygen partial pressure for the cells within minutes, mimicking acute ischemic event. In this study, hypoxia was induced using 0% O gas for three hours and reoxygenation with 19% O gas for 24 hours in serum- and glucose-free medium. According to electrophysiological recordings, hypoxia decreased the hiPSC-CM-beating frequency and field potential (FP) amplitude. Furthermore, FP depolarization time and propagation slowed down. Most of the electrophysiological changes reverted during reoxygenation. However, immunocytochemical staining of the hypoxic and reoxygenation samples showed that morphological changes and changes in the sarcomere structure did not revert during reoxygenation but further deteriorated. qPCR results showed no significant differences in apoptosis or stress-related genes or in the expression of glycolytic genes. In conclusion, the hiPSC-CMs reproduced many characteristic changes of adult CMs during ischemia and reperfusion, indicating their usefulness as a human-based model of acute cardiac ischemia-reperfusion.
缺血性心脏病是最常见的心血管疾病，也是全球医疗保健的主要负担。然而，其病理生理学仍未完全了解，需要基于人体的疾病机制和治疗模型。在此，我们使用人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）在我们新型的细胞培养组件中模拟急性缺血再灌注。该组件能够在数分钟内实现细胞氧分压的交换，模拟急性缺血事件。在本研究中，在无血清和无糖培养基中，使用0%氧气诱导缺氧3小时，然后用19%氧气复氧24小时。根据电生理记录，缺氧降低了hiPSC-CM的搏动频率和场电位（FP）幅度。此外，FP去极化时间和传播速度减慢。大多数电生理变化在复氧过程中恢复。然而，缺氧和复氧样本的免疫细胞化学染色显示，形态学变化和肌节结构变化在复氧过程中并未恢复，反而进一步恶化。qPCR结果显示，凋亡或应激相关基因以及糖酵解基因的表达没有显著差异。总之，hiPSC-CMs在缺血和再灌注过程中重现了成年心肌细胞的许多特征性变化，表明它们作为急性心脏缺血再灌注的基于人体的模型具有实用性。
