Measuring Cardiomyocyte Contractility and Calcium Handling In Vitro
Measuring Cardiomyocyte Contractility and Calcium Handling In Vitro
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DOI：
10.1007/978-1-4939-8597-5_7
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Oh, Jae Gyun
中科院分区：
文献类型：
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作者：
Gorski, Przemek A.;Kho, Changwon;Oh, Jae Gyun
心肌细胞收缩性和 Ca2+ 处理的体外测量已被用作确定各种基因操作的生理后果和确定治疗心力衰竭的潜在治疗靶点的平台。肌细胞钙和收缩系统 (IonOptix) 可同时跟踪单个心肌细胞中的肌节运动和细胞内 Ca2+ 水平的变化。在此，我们描述了一种用于从小鼠心脏中分离成年心肌细胞的修改方案，并提供了如何分析使用 IonOptix 系统收集的心肌细胞 Ca2+ 瞬态和收缩性数据的分步描述。在我们修改后的方案中，我们推荐一种新颖的插管技术，该技术简化了这种困难的方法，并提高了分离心肌细胞的活力。此外，还对细胞内 Ca2+ 处理、SR Ca2+ 负荷、肌丝 Ca2+ 敏感性和心肌细胞收缩力进行了全面分析，以便为心肌力学提供重要见解。
In vitro measurements of cardiomyocyte contractility and Ca2+ handling have been used as a platform for determining physiological consequence of various genetic manipulations and identifying potential therapeutic targets for the treatment of heart failure. The Myocyte Calcium and Contractility System (IonOptix) offers a simultaneous trace of sarcomere movements and changes of intracellular Ca2+ levels in a single cardiomyocyte. Herein, we describe a modified protocol for the isolation of adult cardiomyocytes from murine hearts and provide a step-by-step description on how to analyze cardiomyocyte Ca2+ transient and contractility data collected using the IonOptix system. In our modified protocol, we recommend a novel cannulation technique which simplifies this difficult method and leads to improved viability of isolated cardiomyocytes. In addition, a comprehensive analysis of intracellular Ca2+ handling, SR Ca2+ load, myofilament Ca2+ sensitivity, and cardiomyocyte contractility is described in order to provide important insights into myocardial mechanics.