Department of Medical Biology, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands ; and.
Department of Experimental Cardiology, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands .
J Cardiovasc Pharmacol. 2021 Mar 1;77(3):267-279. doi: 10.1097/FJC.0000000000000955.
Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are supposed to be a good human-based model, with virtually unlimited cell source, for studies on mechanisms underlying cardiac development and cardiac diseases, and for identification of drug targets. However, a major drawback of hPSC-CMs as a model system, especially for electrophysiological studies, is their depolarized state and associated spontaneous electrical activity. Various approaches are used to overcome this drawback, including the injection of "synthetic" inward rectifier potassium current (IK1), which is computed in real time, based on the recorded membrane potential ("dynamic clamp"). Such injection of an IK1-like current results in quiescent hPSC-CMs with a nondepolarized resting potential that show "adult-like" action potentials on stimulation, with functional availability of the most important ion channels involved in cardiac electrophysiology. These days, dynamic clamp has become a widely appreciated electrophysiological tool. However, setting up a dynamic clamp system can still be laborious and difficult, both because of the required hardware and the implementation of the dedicated software. In the present review, we first summarize the potential mechanisms underlying the depolarized state of hPSC-CMs and the functional consequences of this depolarized state. Next, we explain how an existing manual patch clamp setup can be extended with dynamic clamp. Finally, we shortly validate the extended setup with atrial-like and ventricular-like hPSC-CMs. We feel that dynamic clamp is a highly valuable tool in the field of cellular electrophysiological studies on hPSC-CMs and hope that our directions for setting up such dynamic clamp system may prove helpful.
人多能干细胞衍生的心肌细胞（hPSC-CMs）被认为是一种很好的基于人类的模型，具有几乎无限的细胞来源，可用于研究心脏发育和心脏疾病的机制，并用于鉴定药物靶点。然而，hPSC-CMs 作为模型系统的一个主要缺点，特别是对于电生理研究，是其去极化状态和相关的自发电活动。为了克服这一缺点，人们采用了各种方法，包括注入“合成”内向整流钾电流（IK1），该电流是根据记录的膜电位实时计算的（“动态钳位”）。这种 IK1 样电流的注入导致静息电位去极化的静止 hPSC-CMs 表现出“成人样”动作电位，刺激时涉及心脏电生理的最重要离子通道具有功能可用性。如今，动态钳位已成为一种广泛应用的电生理工具。然而，建立动态钳位系统仍然可能是费力和困难的，这既是因为所需的硬件，也是因为专用软件的实现。在本综述中，我们首先总结了 hPSC-CMs 去极化状态的潜在机制及其去极化状态的功能后果。接下来，我们解释了如何用现有的手动膜片钳设置来扩展动态钳位。最后，我们用心房样和心室样 hPSC-CMs 简要验证了扩展设置。我们认为，动态钳位在 hPSC-CMs 的细胞电生理研究领域是一种非常有价值的工具，希望我们建立这种动态钳位系统的指导方向能有所帮助。
