中文摘要：

目的：对新生大鼠心肌细胞分离培养方法进行改进，以期获得较高纯度和活力的心肌细胞；方法：采用胶原酶消化法分离大鼠心肌细胞，控制消化时间、消化温度、搅拌速度、离心次数和速度等，结合贴壁分离和Brd U干预纯化心肌细胞，流式细胞仪分析肌钙蛋白Ⅰ（troponin Ⅰ，Tn Ⅰ）表达，荧光探针进行胞浆游离钙离子染色；结果：多数心肌细胞自动收缩，逐渐聚集形成肌管，troponin Ⅰ阳性率达92％，所有心肌细胞钙离子染色阳性，并随细胞收缩呈周期性变化；结论：该方法分离培养的心肌细胞具有较高的纯度和活力。

英文摘要：

Objective：To improve the isolation and primary culture of newborn rat cardiomyocytes so as to obtain cardiac myocytes with higher purity and vigor. Methods ：Rat cardiomyocytes were isolated with collagenase digestion;the digesting time and temperature ,stirring speed, centrifuging frequency and speed were all under control. Cardiomyocytes were purified by adherence separation and BrdU treatment. Troponin I expression in cardiomyocytes was tested with flow cytometry .Free Ca^2＋ in cytoplasm of cardiomyocytes was detected with fluorescent probe.Results ：Most of the ceils contracted spontaneously, and grew into cluster with the formation of myotube. Positive rate of troponin I staining was 92%, Ca^2＋ fluorescent probe positively expressed in all cardiomyocytes and cycling variation was detected along with cell contraction. Conclusion：Cardiomyocytes with high purity and vigor were got by this way.