In Vivo Assessment of the Electrophysiological Integration and Arrhythmogenic Risk of Myocardial Cell Transplantation Strategies
In Vivo Assessment of the Electrophysiological Integration and Arrhythmogenic Risk of Myocardial Cell Transplantation Strategies
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DOI：
10.1002/stem.545
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发表时间：
2010-12-01
期刊：
影响因子：
5.2
通讯作者：
Olgin, Jeffery E.
中科院分区：
文献类型：
--
作者：
Gepstein, Lior;Ding, Chunhua;Olgin, Jeffery E.
细胞替代策略是旨在改善心肌性能的有前途的干预措施。然而，这些方法的电生理影响尚未阐明。我们评估了两种候选细胞类型，即骨骼肌成肌细胞和人胚胎干细胞衍生的心肌细胞（hESC-CM）移植的电生理后果。将荧光标记（DiO）的候选细胞移植到大鼠左心室心肌中。两周后，使用Langendorff灌注的大鼠心脏制备进行光学标测。用适当的滤波器获得图像，以描绘心脏的解剖结构，识别DiO标记的细胞，并将此信息与电压映射数据（使用电压敏感染料PGH-I）相关联。组织学检查显示移植的骨骼肌肌管和宿主心肌细胞之间缺乏缝隙连接。相反，在hESC-CM移植的心脏中，在供体和宿主心肌细胞之间观察到阳性Cx43免疫染色。光学标测显示hESC-CM植入部位的正常传导（6个中的4个）或最小传导减慢（6个中的2个）。相反，在成肌细胞移植部位观察到明显的传导减慢或传导阻滞（8例中有7例）。程序电刺激后，hESC-CM心脏不能诱导室性心律失常，但在50%的成肌细胞移植心脏中可诱导室性心律失常。总之，提出了一种评估心肌细胞治疗的电生理影响的独特方法。我们的结果证明了hESC-CM与宿主组织功能整合的能力。相反，移植不形成缝隙连接的细胞（骨骼肌成肌细胞）导致局部传导障碍和促神经生长底物的产生。干细胞2010; 28：2151-2161
Cell replacement strategies are promising interventions aiming to improve myocardial performance. Yet, the electrophysiological impact of these approaches has not been elucidated. We assessed the electrophysiological consequences of grafting of two candidate cell types, that is, skeletal myoblasts and human embryonic stem cell-derived cardiomyocytes (hESC-CMs). The fluorescently labeled (DiO) candidate cells were grafted into the rat's left ventricular myocardium. Two weeks later, optical mapping was performed using the Langendorff-perfused rat heart preparation. Images were obtained with appropriate filters to delineate the heart's anatomy, to identify the DiO-labeled cells, and to associate this information with the voltage-mapping data (using the voltage-sensitive dye PGH-I). Histological examination revealed the lack of gap junctions between grafted skeletal myotubes and host cardiomyocytes. In contrast, positive Cx43 immunostaining was observed between donor and host cardiomyocytes in the hESC-CMs-transplanted hearts. Optical mapping demonstrated either normal conduction (four of six) or minimal conduction slowing (two of six) at the hESC-CMs engraftment sites. In contrast, marked slowing of conduction or conduction block was seen (seven of eight) at the myoblast transplantation sites. Ventricular arrhythmias could not be induced in the hESC-CM hearts following programmed electrical stimulation but were inducible in 50% of the myoblast-engrafted hearts. In summary, a unique method for assessment of the electrophysiological impact of myocardial cell therapy is presented. Our results demonstrate the ability of hESC-CMs to functionally integrate with host tissue. In contrast, transplantation of cells that do not form gap junctions (skeletal myoblats) led to localized conduction disturbances and to the generation of a proarrhythmogenic substrate. STEM CELLS 2010; 28: 2151-2161