Heart Science Centre, National Heart & Lung Institute, Imperial College London, UK.
J Mol Cell Cardiol. 2011 Sep;51(3):390-8. doi: 10.1016/j.yjmcc.2011.06.018. Epub 2011 Jun 29.
Electrophysiological and pharmacological data from the human heart are limited due to the absence of simple but representative experimental model systems of human myocardium. The aim of this study was to establish and characterise adult human myocardial slices from small patients' heart biopsies as a simple, reproducible and relevant preparation suitable for the study of human cardiac tissue at the multicellular level. Vibratome-cut myocardial slices were prepared from left ventricular biopsies obtained from end-stage heart failure patients undergoing heart transplant or ventricular assist device implantation, and from hearts of normal dogs. Multiple slices were prepared from each biopsy. Regular contractility was observed at a range of stimulation frequencies (0.1-2 Hz), and stable electrical activity, monitored using multi-electrode arrays (MEA), was maintained for at least 8 h from slice preparation. ATP/ADP and phosphocreatine/creatine ratios were comparable to intact organ values, and morphology and gap junction distribution were representative of native myocardium. MEA recordings showed that field potential duration (FPD) and conduction velocity (CV) in human and dog slices were similar to the values previously reported for papillary muscles, ventricular wedges and whole hearts. Longitudinal CV was significantly faster than transversal CV, with an anisotropic ratio of 3:1 for human and 2.3:1 for dog slices. Importantly, slices responded to the application of E-4031, chromanol and 4-aminopyridine, three potassium channel blockers known to affect action potential duration, with an increase in FPD. We conclude that viable myocardial slices with preserved structural, biochemical and electrophysiological properties can be prepared from adult human and canine heart biopsies and offer a novel preparation suitable for the study of heart failure and drug screening.
由于缺乏简单但具有代表性的人类心肌实验模型系统,人类心脏的电生理和药理学数据有限。本研究的目的是建立和表征成人心肌切片,这些切片来自于小患者的心脏活检,作为一种简单、可重复和相关的准备,适用于在多细胞水平上研究人类心脏组织。使用振动切片机从终末期心力衰竭患者接受心脏移植或心室辅助装置植入术的左心室活检以及正常犬的心脏中制备心肌切片。从每个活检中制备多个切片。在 0.1-2Hz 的刺激频率范围内观察到规则的收缩性,并且使用多电极阵列(MEA)监测的稳定电活动至少在切片制备 8 小时内保持稳定。ATP/ADP 和磷酸肌酸/肌酸比与完整器官的值相当,形态和缝隙连接分布代表了原生心肌。MEA 记录显示,人类和犬切片的场电位持续时间(FPD)和传导速度(CV)与先前报道的乳头肌、心室楔形和整个心脏的数值相似。纵向 CV 明显快于横向 CV,人类和犬切片的各向异性比分别为 3:1 和 2.3:1。重要的是,切片对 E-4031、chromanol 和 4-氨基吡啶的应用有反应,这三种钾通道阻滞剂已知会影响动作电位持续时间,导致 FPD 增加。我们得出结论,从成人人类和犬心脏活检中可以制备具有保留结构、生化和电生理特性的可行心肌切片,为心力衰竭和药物筛选的研究提供了一种新的准备方法。