Methods for assessing the electromechanical integration of human pluripotent stem cell-derived cardiomyocyte grafts.
Methods for assessing the electromechanical integration of human pluripotent stem cell-derived cardiomyocyte grafts.
复制标题
评估人多能干细胞来源的心肌细胞移植物机电一体化的方法。
DOI：
10.1007/978-1-4939-1047-2_20
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Laflamme,MichaelA
中科院分区：
文献类型：
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作者：
Zhu,Wei-Zhong;Filice,Dominic;Palpant,NathanJ;Laflamme,MichaelA
从人多能干细胞中提取的心肌细胞在替代心肌和心肌收缩功能方面表现出巨大的潜力。然而，直到最近，还没有方法可以直接确定这些干细胞来源的移植物是否在移植后与宿主心肌同步结合，并在完整或受伤的心脏中同步燃烧。为了解决这一不确定性，我们的研究小组开发了一种技术，用于对移植了干细胞来源的心肌细胞的心脏进行活体成像，这些心肌细胞已经被修饰为表达基因编码的蛋白质钙传感器GCaMP。当与同时记录的心电图相结合时，该方案允许人们对宿主-移植物电耦合的存在和程度以及移植物激活的时间和模式进行定量评估。如本文所述，该系统已被用于研究人类胚胎干细胞来源的心肌细胞在豚鼠心脏损伤模型中的机电一体化，但类似的方法应适用于其他人类移植细胞类型和动物模型。
Cardiomyocytes derived from human pluripotent stem cells show tremendous promise for the replacement of myocardium and contractile function lost to infarction. However, until recently, no methods were available to directly determine whether these stem cell-derived grafts actually couple with host myocardium and fire synchronously following transplantation in either intact or injured hearts. To resolve this uncertainty, our group has developed techniques for the intravital imaging of hearts engrafted with stem cell-derived cardiomyocytes that have been modified to express the genetically encoded protein calcium sensor, GCaMP. When combined with the simultaneously recorded electrocardiogram, this protocol allows one to make quantitative assessments as to the presence and extent of host–graft electrical coupling as well as the timing and pattern of graft activation. As described here, this system has been employed to investigate the electromechanical integration of human embryonic stem cell-derived cardiomyocytes in a guinea pig model of cardiac injury, but analogous approaches should be applicable to other human graft cell types and animal models.