Objective:To introduce an emergent isolation method for acquiring adult rats myocytes stably to favor the study on myocardial electrophysiology and molecular biology. Methods:The heart of Sprague-Dawley(SD) adult rat was rapidly excised after successful anesthesia and mounted on a Langendorff apparatus .It was perfused retrogradely via the aorta for five minutes with Ca2+-free Tyrode’s solution

and then perfused with collagenase dissolved in nominally Ca2+-free Tyrode’s solution for twenty minutes

then the rat ventricular cells were isolated by seperate times and stored in a Kraftbruhe(KB) solution. Results:A large amount of rat ventricular cells in high activity(78%±7.5%)were acquired

they were in good shaps and their cross-veins were clear. Conclusion:The introduced isolation method is with good stability and can acquire a large mount of myocardial cells in high activity

which are applicable for the study on myocardial electrophsiology and molecular biology.