新生SD大鼠心肌细胞的原代培养及荧光鉴定  

Primary culture and identification of cardiomyocytes of neonatal SD rats

作　　者：姬婷婷[1] 徐岩[1] 张建华[1] 赵勇[1] 

机构地区：[1]安徽医科大学第一附属医院心血管内科,合肥230022

出　　处：《安徽医科大学学报》2014年第1期120-122,共3页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:040713103)

摘　　要：采用单纯胰蛋白酶消化法分离,差速贴壁法纯化新生大鼠心肌细胞。光学显微镜观察心肌细胞基本形态特征,台盼蓝染色检测成活率,肌钙蛋白Ⅰ、α-横纹肌肌动蛋白抗体鉴定心肌细胞。结果显示培养24 h后心肌细胞大部分已贴壁,并出现自发搏动现象,48 h后逐渐伸出伪足相互接触交织成网,72 h后逐渐形成细胞簇并出现同步搏动;台盼蓝染色法检测细胞成活率达90%以上;肌钙蛋白Ⅰ及α-横纹肌肌动蛋白免疫荧光染色法鉴定心肌细胞纯度达98%以上。Neonatal rat myocytes were digested with typsin and purified by technique of differential anchoring. The cardiomyocytes cultured were observed under light microscope, stained with trypan blue to count the living cells and identified with cTnI and α-Sarcomeric aetin. The results showed most of cardiomyocytes adherented to culture plates after 24 hours of culture and a part of cells were jumping synchronously, 48 hours later cells gradually extended pseudopodia, contacted with each other and interwovened into a network. After 72 hours of culture, the ceils gradually formed cell clusters and were beating synchronously. Trypan blue staining showed the survival rate of cardiomyocytes was more than 90%. cTnI and α-Sarcomeric actin immunofluorescence staining showed that the purity of cardiomyocytes cultured was more than 98%.

关 键 词：心肌细胞 原代培养 新生大鼠 免疫荧光 

分 类 号：R540.49[医药卫生—心血管疾病]