Coppini Raffaele, Ferrantini Cecila, Aiazzi Alessandro, Mazzoni Luca, Sartiani Laura, Mugelli Alessandro, Poggesi Corrado, Cerbai Elisabetta
Department NeuroFarBa, Division of Pharmacology, University of Florence;
Department of Clinical and Experimental Medicine, Division of Physiology, University of Florence.
J Vis Exp. 2014 Apr 21(86):51116. doi: 10.3791/51116.
Cardiomyocytes from diseased hearts are subjected to complex remodeling processes involving changes in cell structure, excitation contraction coupling and membrane ion currents. Those changes are likely to be responsible for the increased arrhythmogenic risk and the contractile alterations leading to systolic and diastolic dysfunction in cardiac patients. However, most information on the alterations of myocyte function in cardiac diseases has come from animal models. Here we describe and validate a protocol to isolate viable myocytes from small surgical samples of ventricular myocardium from patients undergoing cardiac surgery operations. The protocol is described in detail. Electrophysiological and intracellular calcium measurements are reported to demonstrate the feasibility of a number of single cell measurements in human ventricular cardiomyocytes obtained with this method. The protocol reported here can be useful for future investigations of the cellular and molecular basis of functional alterations of the human heart in the presence of different cardiac diseases. Further, this method can be used to identify novel therapeutic targets at cellular level and to test the effectiveness of new compounds on human cardiomyocytes, with direct translational value.
来自患病心脏的心肌细胞会经历复杂的重塑过程，包括细胞结构、兴奋收缩偶联和膜离子电流的变化。这些变化可能是导致心律失常风险增加以及收缩和舒张功能障碍的收缩改变的原因，而收缩和舒张功能障碍会出现在心脏病患者中。然而，关于心脏病中心肌细胞功能改变的大多数信息都来自动物模型。在此，我们描述并验证了一种从接受心脏手术的患者的心室心肌小手术样本中分离存活心肌细胞的方案。该方案将详细描述。据报道，通过电生理和细胞内钙测量可证明用该方法获得的人心室心肌细胞中许多单细胞测量的可行性。本文报道的方案可能有助于未来对不同心脏病情况下人类心脏功能改变的细胞和分子基础进行研究。此外，该方法可用于在细胞水平识别新的治疗靶点，并测试新化合物对人心肌细胞的有效性，具有直接的转化价值。
