Progressive maturation in contracting cardiomyocytes derived from human embryonic stem cells: Qualitative effects on electrophysiological responses to drugs
Progressive maturation in contracting cardiomyocytes derived from human embryonic stem cells: Qualitative effects on electrophysiological responses to drugs
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DOI：
10.1016/j.scr.2010.01.002
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发表时间：
2010-05-01
影响因子：
1.2
通讯作者：
Nakatsuji, Norio
中科院分区：
文献类型：
--
作者：
Otsuji, Tomomi G.;Minami, Itsunari;Nakatsuji, Norio
药物检测领域目前需要一种新的集成分析系统，与使用天然组织的系统一样准确，这将使我们能够预测候选药物的心律失常风险以及基因突变与获得的电生理表型之间的关系。这可以通过将微电极阵列（MEA）系统与来自人胚胎干细胞（hESC）和诱导多能干细胞的心肌细胞（CM）相结合来实现。CM已经成功地从两种类型诱导，但它们的成熟过程没有系统地控制;这导致搏动效力的丧失和离子通道功能不足。我们产生了转基因hESC系，其通过表达由心脏特异性α MHC启动子驱动的GFP，每2周促进hESC-CM簇的维持，从而在一年内在心室群体内产生紧凑的起搏器谱系。进一步的分析，包括定量RT-PCR，膜片钳，和MEA介导的QT测试，表明，重新铺板培养不断增强基因表达，离子电流幅度，并在hESC-CM的K+通道阻滞的阻力。此外，时间三维（3D）培养加速成熟恢复全球基因抑制状态建立在粘附状态。因此，再铺板/3D培养产生了hESC-CM，其充当适用于再生医学和精确药物测试的功能性合胞体。(C)2010爱思唯尔有限公司版权所有。
The field of drug testing currently needs a new integrated assay system, as accurate as systems using native tissues, that will allow us to predict arrhythmia risks of candidate drugs and the relationship between genetic mutations and acquired electrophysiological phenotypes. This could be accomplished by combining the microelectrode array (MEA) system with cardiomyocytes (CMs) derived from human embryonic stem cells (hESC) and induced pluripotential stem cells. CMs have been successfully induced from both types, but their maturation process is not systematically controlled; this results in loss of beating potency and insufficient ion channel function. We generated a transgenic hESC line that facilitates maintenance of hESC-CM clusters every 2 weeks by expressing GFP driven by a cardiac-specific alpha MHC promoter, thereby producing a compact pacemaker lineage within a ventricular population over a year. Further analyses, including quantitative RT-PCR, patch-clamp, and MEA-mediated QT tests, demonstrated that replating culturing continuously enhanced gene expression, ionic current amplitudes, and resistance to K+ channel blockades in hESC-CMs. Moreover, temporal three-dimensional (3D) culturing accelerated maturation by restoring the global gene repressive status established in the adhesive status. Replating/3D culturing thus produces hESC-CMs that act as functional syncytia suitable for use in regenerative medicine and accurate drug tests. (C) 2010 Elsevier B.V. All rights reserved.