Kijlstra Jan David, Hu Dongjian, van der Meer Peter, Domian Ibrahim J
Cardiovascular Research Center, Massachusetts General Hospital, Boston, Massachusetts.
Harvard Medical School, Boston, Massachusetts.
Curr Protoc Stem Cell Biol. 2017 Nov 15;43:1F.20.1-1F.20.9. doi: 10.1002/cpsc.40.
Human pluripotent stem-cell derived cardiomyocytes (hPSC-CMs) hold great promise for applications in human disease modeling, drug discovery, cardiotoxicity screening, and, ultimately, regenerative medicine. The ability to study multiple parameters of hPSC-CM function, such as contractile and electrical activity, calcium cycling, and force generation, is therefore of paramount importance. hPSC-CMs cultured on stiff substrates like glass or polystyrene do not have the ability to shorten during contraction, making them less suitable for the study of hPSC-CM contractile function. Other approaches require highly specialized hardware and are difficult to reproduce. Here we describe a protocol for the preparation of hPSC-CMs on soft substrates that enable shortening, and subsequently the simultaneous quantitative analysis of their contractile and electrical activity, calcium cycling, and force generation at single-cell resolution. This protocol requires only affordable and readily available materials and works with standard imaging hardware. © 2017 by John Wiley & Sons, Inc.
人多能干细胞衍生的心肌细胞（hPSC-CMs）在人类疾病建模、药物发现、心脏毒性筛查以及最终的再生医学应用中具有巨大潜力。因此，研究hPSC-CM功能的多个参数，如收缩和电活动、钙循环以及力的产生，能力至关重要。在玻璃或聚苯乙烯等坚硬基质上培养的hPSC-CMs在收缩时没有缩短的能力，这使得它们不太适合用于研究hPSC-CM的收缩功能。其他方法需要高度专业化的硬件且难以重复。在这里，我们描述了一种在软基质上制备hPSC-CMs的方案，该方案能够实现缩短，随后以单细胞分辨率对其收缩和电活动、钙循环以及力的产生进行同步定量分析。该方案仅需要价格合理且易于获得的材料，并可与标准成像硬件配合使用。© 2017约翰威立父子公司。
