冷冻保存对人诱导多能干细胞衍生心肌细胞评估药物安全性反应特征的影响。

Effects of Cryopreservation on Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes for Assessing Drug Safety Response Profiles.

机构信息

Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA; Division of Cardiovascular Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA; Molecular Imaging Program at Stanford, Department of Radiology, Stanford University School of Medicine, Stanford, CA, USA.

出版信息

Stem Cell Reports. 2021 Jan 12;16(1):168-181. doi: 10.1016/j.stemcr.2020.11.010. Epub 2020 Dec 17.

Burgeoning applications of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in disease modeling, regenerative medicine, and drug screening have broadened the usage of hiPSC-CMs and entailed their long-term storage. Cryopreservation is the most common approach to store hiPSC-CMs. However, the effects of cryopreservation and recovery on hiPSC-CMs remain poorly understood. Here, we characterized the transcriptome, electro-mechanical function, and drug response of fresh hiPSC-CMs without cryopreservation and recovered hiPSC-CMs from cryopreservation. We found that recovered hiPSC-CMs showed upregulation of cell cycle genes, similar or reduced contractility, Ca transients, and field potential duration. When subjected to treatment of drugs that affect electrophysiological properties, recovered hiPSC-CMs showed an altered drug response and enhanced propensity for drug-induced cardiac arrhythmic events. In conclusion, fresh and recovered hiPSC-CMs do not always show comparable molecular and physiological properties. When cryopreserved hiPSC-CMs are used for assessing drug-induced cardiac liabilities, the altered drug sensitivity needs to be considered.

新兴的人类诱导多能干细胞衍生心肌细胞（hiPSC-CMs）在疾病建模、再生医学和药物筛选中的应用拓宽了 hiPSC-CMs 的用途，并需要对其进行长期储存。冷冻保存是储存 hiPSC-CMs 的最常用方法。然而，冷冻保存及其恢复对 hiPSC-CMs 的影响仍知之甚少。在这里，我们对未经冷冻保存的新鲜 hiPSC-CMs 和从冷冻保存中恢复的 hiPSC-CMs 的转录组、机电功能和药物反应进行了表征。我们发现，恢复的 hiPSC-CMs 中细胞周期基因上调，收缩力、Ca 瞬变和场电位持续时间相似或降低。当用影响电生理特性的药物处理时，恢复的 hiPSC-CMs 表现出改变的药物反应和增强的药物诱导心律失常事件的倾向。总之，新鲜和恢复的 hiPSC-CMs 并不总是表现出相似的分子和生理特性。当使用冷冻保存的 hiPSC-CMs 来评估药物引起的心脏毒性时，需要考虑改变的药物敏感性。