BACKGROUND: To obtain high survival rate and good activity of the normal single myocardial cells is the basis of myocardialelectrophysiology. It is important to find an experimental method which is close to the normal myocardial cells in morphology,function and structure.OBJECTIVE: To discuss the influence factors during emergent isolation of myocardial cells by using enzymatic method in ratsand to build an emergent isolation method for stable obtaining myocardial cells to provide cells for myocardial electrophysiologyand molecular biology.METHODS: The heart of Sprague-Dawley adult rats was rapidly excised after cervical dislocated and mounted on a Langendorffapparatus for aortic counter-current perfusion. It was perfused retrograde via the aorta for 5 minutes with calcium-free tyrode, andthen perfused with collagenase dissolved in nominally calcium-free tyrode for 20 minutes. Ultimately calcium solution wasperfused for 3 minutes. Then the rat ventricular cells were stored in HK solution.RESULTS AND CONCLUSION: Viability of freshly isolated myocardial cells which were rod-shaped with clear-striation was60%-80%. It is shown that to adjust and control all the factors in the emergent isolation can ensure the quantity and quality ofmyocardial cells.%背景:获得存活率高、活性好的正常单个心肌细胞是心肌电生理研究的基础,寻求一种可以获得在形态、功能和结构上接近于正常心肌细胞的实验方法具有重要意义.目的:探讨酶解法急性分离大鼠心肌细胞过程中的各影响因素,建立一种可以稳定获取高活性心肌细胞的急性分离方法,为心肌细胞电生理研究和分子生物学研究提供细胞.方法:SD 大鼠颈椎脱臼,迅速开胸取心脏,将心脏悬挂于Langendorff 装置上进行主动脉逆流灌流.先用无钙台式液灌流5 min,然后换上酶液灌流消化20 min,最后用终止液灌流3 min 终止反应,将细胞置于HK 液中保存.结果与结论:可获得60%~80%的横纹清晰,透亮的长杆状的心肌细胞.说明调节和控制细胞分离过程中的各影响因素可以保证分离的心肌细胞的数量和质量.
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