Single-Cell Functional Analysis of Stem-Cell Derived Cardiomyocytes on Micropatterned Flexible Substrates.
Single-Cell Functional Analysis of Stem-Cell Derived Cardiomyocytes on Micropatterned Flexible Substrates.
复制标题
DOI：
10.1002/cpsc.40
复制
发表时间：
2017-11-15
影响因子：
--
通讯作者：
Domian, Ibrahim J
中科院分区：
文献类型：
--
作者：
Kijlstra, Jan David;Hu, Dongjian;Domian, Ibrahim J
人类多能干细胞来源的心肌细胞（hPSC-CMs）在人类疾病建模、药物发现、心脏毒性筛选以及最终的再生医学方面有着巨大的应用前景。因此，研究hPSC-CM功能的多个参数，如收缩和电活动、钙循环和力产生的能力是至关重要的。在玻璃或聚苯乙烯等坚硬基质上培养的hspc - cm在收缩过程中不具有缩短的能力，因此不太适合研究hspc - cm的收缩功能。其他方法需要高度专业化的硬件，而且很难复制。在这里，我们描述了一种在软底物上制备hPSC-CMs的方案，该方案可以缩短，随后在单细胞分辨率下同时定量分析其收缩和电活动，钙循环和力产生。该协议只需要负担得起且容易获得的材料，并与标准成像硬件一起工作。©2017 by John Wiley & Sons, Inc。
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.