酶消化法分离培养大鼠血管平滑肌细胞方法的改良  

Improvememt of the methods for culture of rat vascular smooth muscle cells by enzymatic digestion

作　　者：李宪伟[1] 姜维良[1] 

机构地区：[1]哈尔滨医科大学附属第二医院血管外科,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2011年第1期58-60,63,共4页Journal of Harbin Medical University

基　　金：黑龙江省教育厅骨干教师项目(1153G018)

摘　　要：目的改良酶消化法并建立一种简单方便且高效的血管平滑肌细胞原代培养方法。方法在无菌条件下分离大鼠胸主动脉,0.1%Ⅱ型胶原酶消化分离血管平滑肌细胞并进行培养。用倒置相差显微镜观察血管平滑肌细胞的生物学特性,用免疫组化染色法检测细胞胞浆内α-肌动蛋白(α-actin)的表达。结果相差显微镜下细胞呈现"谷和峰"的生长特点,α-actin胞浆染色阳性细胞数占总细胞数的98%以上。结论单纯Ⅱ型胶原酶可消化分离培养血管平滑肌细胞,且有成活细胞数多、传代周期短的特点。Objective To reform enzymatic digestion and create a simple and effective primary culture methods for rat vascular smooth muscle cells（VSMC）.Methods 0.1% collagenase Ⅱ was used to digest and isolate the thoracic aorta smooth muscle cells of rat and then cultured.The feature of vascular smooth muscle cells was observed by inverted phase contrast microscope and observed α-actin stained by indirect immunocytochemistry.Results The cultured cells exhibited the characteristic ＂hills and valleys＂ growth pattern as observed by phase contrast microscopy and cells that showed α-SM-actin positive staining were more than 98%.Conclusion Collagenase Ⅱ can digest and isolate vascular smooth muscle cells and also has advantages that make cells easy to live and short period for passaging.

关 键 词：胸主动脉 血管平滑肌细胞 大鼠 培养 

分 类 号：R-332[医药卫生]