酶消化法急性分离小鼠颈总动脉平滑肌细胞实验研究  

Enzymatic Isolation of Common Carotid Artery Vascular Smooth Muscle Cells of Mouse

作　　者：苏现秀 丁良[1] 

机构地区：[1]河北大学基础医学院,河北保定071000

出　　处：《亚太传统医药》2016年第4期11-13,共3页Asia-Pacific Traditional Medicine

摘　　要：目的:探讨快速分离小鼠颈总动脉血管平滑肌细胞(CCAVSMCs)的实验方法。方法:分离小鼠颈总动脉,用酶液Ⅰ(木瓜蛋白酶等)、酶液Ⅱ(Ⅱ型胶原酶)消化获取平滑肌细胞,并确定两步反应中酶各自的最佳消化时间,以台盼蓝染色测定细胞活性,采用免疫荧光法进行细胞鉴定。结果:在倒置显微镜下,观察并计数分离的平滑肌细胞数,酶液Ⅰ、酶液Ⅱ的消化时间分别为25min、45min时平滑肌细胞数量较多,达(50±1.3)个/低倍视野,与其他各时间点比较差异具有统计学意义(P<0.05),98%的台盼蓝染色细胞为活细胞,免疫荧光法鉴定为平滑肌细胞。结论:该方法操作简单、成本低,可在较短时间内获取数量可观、形态佳、高纯度的颈总动脉平滑肌细胞。Objective：To investigate a rapidly enzymatic isolation of common carotid artery vascular smooth muscle cells of mouse.Methods：Carotid vascular smooth muscle cells of mouse were isolated.EnzymeⅠ（papain etc）and enzymeⅡ（collagenase typeⅡ）were used to digest carotid vascular smooth muscle cells of mouse.The respective optimal digestion time of these two kinds of enzymes in two steps reaction were dertermined.The cell activity was determined by trypan blue staining.The cells were identified by immunoflourescent technique.Results：The isolated smooth muscle cells were observed and counted under inverted microscope.The optimal digestion time of enzymeⅠand enzymeⅡwas 25 and 45min respectively,the number of isolated smooth muscle cells were（50±1.3）per low power field and significant differences were shown with other digestion time（P〈0.05）.The cells identified by immunoflourescent technique were smooth muscle cells and living cells detected by trypan blue staining were more than 98 percents.Conclusion：The method which has lower cost is easy,by which considerable,good form and high purity of the common carotid artery smooth muscle cells can be obtainted in a short time.

关 键 词：血管平滑肌细胞 酶消化 测定 

分 类 号：R743.7[医药卫生—神经病学与精神病学]