原代心肌细胞培养

目的:探讨SD大鼠乳鼠心肌细胞的分离及培养方法,并进行形态学观察。方法:取出生1～3 d的SD大鼠的乳鼠,胰蛋白酶和Ⅱ型胶原酶分别消化心肌组织,差速贴壁法分离心肌细胞,并加入适量BrdU纯化心肌细胞,显微镜下观察心肌细胞形态及纯度。结果:成功分离培养心肌细胞并观察其形态,计算存活率92%,纯度90%,2～3 d出现同步搏动。结论:胰蛋白酶联合Ⅱ型胶原酶消化可分离存活率较高的心肌细胞,用差速贴壁法并加入适量BrdU可纯化心肌细胞,使细胞搏动良好。

The culture of primary cardiac cells

Objective:To study the primary culture and observe the morphological features of myocardial cells of SD neonatal rats.Methods:The ventricular myocardia of SD neonatal rats with an age of 1~3 days were obtained,and they were digested with trypsin,then digested with collagenase Ⅱ.Myocardial cells were isolated with the technique of differential anchoring velocity with 90 min and were pured by BrdU,then observe the myocardial cells form and purity by microscope.Results:The culture was successful with these methods.The morphological features of myocardial cells was observed.The survival rate was 92% and purity was 90%.The pulsation of all myocardical cells was synchronous.Conclusion:Using trypsin and collagenase Ⅱ with the technique of differential anchoring velocity to isolate and culture myocardial cells can improve survival rate and purity of primary myocardial cells,and enhance cells contraction.