原代心房肌细胞的培养及鉴定

Primary culture and identification of rat atrial myocytes

目的 探讨心房肌细胞原代培养的方法，为对房颤早期电重构的研究奠定基础。方法 采用2周左右的大鼠，取左、右心房，胰蛋白酶结合Ⅱ型胶原酶消化细胞，利用差异性贴壁技术及加用Brdu纯化心房肌细胞，观察细胞形态以及结合免疫细胞化学检测心肌细胞特异性表达的α-肌动蛋白鉴定心房肌细胞。结果 培养至第4天，细胞生长密度可以达到瓶底70％左右，数目可达2～3×10^6／ml，经免疫细胞化学鉴定，90％以上培养细胞a-肌动蛋白抗体染色阳性。结论 利用酶消化法成功培养出大鼠心房肌细胞，所得心房肌细胞纯度高，为进一步深入研究房颤时心房肌细胞的重构机制奠定了基础。

Objective To explore a method for primary culture and identification of rat atrial myocytes. Methods Specimens of rat atria were resected from two weeks old rat. Primary atrial myocytes were cultured by enzyme dissection method（trypsogen and collagenase Ⅱ ）. Atrial myocardial cells were purified by technique of differential platting and adding Brdu and identified by immunocytochemical analysis of α-actin. Results After 4d of primary culture, the number of cells reached （2-3） × 10^6/ml. Immunocytochemical analysis showed that over 90 percent of cells were stained positively for α-actin,a factor specific for cardiomyocytes. Conclusion Rat atrial myocytes were successfully cultured by enzyme dissection method with high purity.