Division of Cardiovascular Sciences, Myocardial Function, National Heart and Lung Institute, Imperial College London, London, UK.
Translational Biomedical Research Centre, University of Bristol, Bristol, UK.
Nat Protoc. 2017 Dec;12(12):2623-2639. doi: 10.1038/nprot.2017.139. Epub 2017 Nov 30.
This protocol describes the preparation of highly viable adult ventricular myocardial slices from the hearts of small and large mammals, including rodents, pigs, dogs and humans. Adult ventricular myocardial slices are 100- to 400-μm-thick slices of living myocardium that retain the native multicellularity, architecture and physiology of the heart. This protocol provides a list of the equipment and reagents required alongside a detailed description of the methodology for heart explantation, tissue preparation, slicing with a vibratome and handling of myocardial slices. Supplementary videos are included to visually demonstrate these steps. A number of critical steps are addressed that must be followed in order to prepare highly viable myocardial slices. These include identification of myocardial fiber direction and fiber alignment within the tissue block, careful temperature control, use of an excitation-contraction uncoupler, optimal vibratome settings and correct handling of myocardial slices. Many aspects of cardiac structure and function can be studied using myocardial slices in vitro. Typical results obtained with hearts from a small mammal (rat) and a large mammal (human) with heart failure are shown, demonstrating myocardial slice viability, maximum contractility, Ca handling and structure. This protocol can be completed in ∼4 h.
本方案描述了从小鼠、猪、犬和人等小型和大型哺乳动物心脏中制备高活力成年心室心肌切片的方法。成年心室心肌切片是厚度为 100-400μm 的活心肌切片，保留了心脏的固有细胞多样性、结构和生理学特性。本方案提供了所需设备和试剂的清单，并详细描述了心脏切片的组织准备、振动切片机切片和心肌切片处理等步骤。此外，还包括了一些视频以直观地展示这些步骤。本方案还介绍了一些关键步骤，必须严格遵循这些步骤才能制备高活力的心肌切片。其中包括识别组织块内的心肌纤维方向和纤维排列、严格控制温度、使用兴奋-收缩偶联抑制剂、优化振动切片机设置以及正确处理心肌切片。使用体外心肌切片可以研究心脏的许多结构和功能方面。展示了从小鼠（大鼠）和患有心力衰竭的大型哺乳动物（人）心脏获得的典型结果，证明了心肌切片的活力、最大收缩性、Ca 处理和结构。本方案大约可在 4 小时内完成。
