Variability of Action Potentials Within and Among Cardiac Cell Clusters Derived from Human Embryonic Stem Cells.
Variability of Action Potentials Within and Among Cardiac Cell Clusters Derived from Human Embryonic Stem Cells.
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DOI：
10.1038/srep18544
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发表时间：
2016-01-05
影响因子：
4.6
通讯作者：
Tung L
中科院分区：
文献类型：
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作者：
Zhu R;Millrod MA;Zambidis ET;Tung L
多能干细胞来源的心肌细胞的电生理变异仍然是其科学和转化应用的障碍。我们使用高分辨率光学测绘研究了从人类胚胎干细胞衍生的心肌细胞 (hESC-CM) 簇记录的动作电位 (AP) 的变异性。通过四个参数分析了超过 23,000 个 AP：APD30、APD80、三角测量和分数复极。尽管采取了措施来减少由于细胞培养条件和 AP 速率依赖性造成的变异性，但我们仍然观察到簇内和簇内 AP 的显着变异性。然而，在非常接近的空间位置中发现了类似的 AP，并且在一些簇中形成了具有不同 AP 特征的不同区域，这些区域在 AP 参数分布中反映为单独的峰，表明存在多种电生理表型。使用最近开发的自动化方法根据整个 AP 形状对细胞进行分组，当将 AP 分为 2 或 3 个亚群时，我们识别出单个簇内不同表型的不同区域以及不同簇之间的常见表型。对大量 hESC-CM 的异质性和潜在表型的系统分析可用于评估提高多能干细胞来源的心肌细胞质量的策略，以用于诊断和治疗应用以及药物筛选。
Electrophysiological variability in cardiomyocytes derived from pluripotent stem cells continues to be an impediment for their scientific and translational applications. We studied the variability of action potentials (APs) recorded from clusters of human embryonic stem cell-derived cardiomyocytes (hESC-CMs) using high-resolution optical mapping. Over 23,000 APs were analyzed through four parameters: APD30, APD80, triangulation and fractional repolarization. Although measures were taken to reduce variability due to cell culture conditions and rate-dependency of APs, we still observed significant variability in APs among and within the clusters. However, similar APs were found in spatial locations with close proximity, and in some clusters formed distinct regions having different AP characteristics that were reflected as separate peaks in the AP parameter distributions, suggesting multiple electrophysiological phenotypes. Using a recently developed automated method to group cells based on their entire AP shape, we identified distinct regions of different phenotypes within single clusters and common phenotypes across different clusters when separating APs into 2 or 3 subpopulations. The systematic analysis of the heterogeneity and potential phenotypes of large populations of hESC-CMs can be used to evaluate strategies to improve the quality of pluripotent stem cell-derived cardiomyocytes for use in diagnostic and therapeutic applications and in drug screening.