大鼠血管平滑肌细胞的培养与鉴定

Culture and Identification of Rat Vascular Smooth Muscle Cells

目的:改进大鼠血管平滑肌细胞(VSMCs)的培养方法。方法:采用机械刮除、差异贴壁等手段进行组织贴块法原代培养,胰蛋白酶消化传代,并应用相差显微镜和即用型SABC免疫组化染色试剂盒对细胞进行形态学和免疫组化鉴定。结果:镜下培养细胞呈典型的"谷-峰状"生长,免疫组化染色显示胞浆内-αactin阳性表达,锥虫蓝染色检测细胞传代成活率95%。结论:本法可获纯度高、结构和功能良好的VSMCs。

Objective： To improve the culture method of vascular smooth muscle cells（VSMCs）in rats. Methods： The primary culture and subculture were done by modified tissue-piece inoculation and trypsin digestion respectively. The cells were purified by the mechanical treatment and differential attachment. The cultured ceils were identified by morphology and immunohistochemistry with phase contrast microscope and SABC kit. Results： The cultured cells possessed ＂peak and valley＂ characteristics for VSMCs under microscope. Immunohistochemical staining with antibody against SM- α-actin demonstrated these ceils were positive. The cell viability was 95% in subculture by trypan blue. Conclusion： The highly purified VSMCs with good structure and function are obtained by this method.