乳鼠脑血管平滑肌细胞的分离培养与鉴定  

Isolating culture and identification of cerebraovascular smooth muscle cells from neonatal mice

作　　者：李世[1] 侯雪芹[1] 陈云波[1] 程淑意[1] 王奇[1] 

机构地区：[1]广州中医药大学临床药理研究所/DME中心,510405

出　　处：《中华老年心脑血管病杂志》2014年第12期1316-1318,共3页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(81273817)

摘　　要：目的建立一种快速、简单、高效的脑血管平滑肌细胞分离和培养方法,对脑血管疾病机制的研究提供实验材料基础。方法用不含EDTA胰蛋白酶消化组织获取细胞,继而于倒置相差显微镜下观察细胞的生长情况,对第3代细胞采用α-肌动蛋白荧光免疫染色法,同时MTT检测细胞的生长趋势。结果运用胰酶消化法获得并培养出大量的脑血管平滑肌细胞,荧光免疫染色法鉴定结果显示阳性表达,纯度98%。结论此方法简单、操作性强,目标细胞获得率高,为老年性脑血管疾病机制研究提供材料支持。Objective To study the pathogenesis of cerebraovascular diseases by establishing a rapid,simple and effective isolation and culture method for cerebrovascular smooth muscle cells from neonatal mice.Methods Cerebrovascular smooth muscle cells were harvested from the trypsinized tissue of neonatal mice containing no EDTA.Their growth was observed under inverted phase contrast microscope and assayed by MTT assay with alpha-actin immunofluorsence staining.Results Massive cerebrovascular smooth muscle cells were harvested and cultured using trypsin digestion technique with a purity of 98％.Their positive expression was identified with immunofluorsence staining.Conclusion The method we established in this study can harvest massive target cells due to its simple and easy operability,thus providing a material support for studying the pathogenesis of cerebrovascular diseases in the elderly.

关 键 词：脑血管障碍 肌细胞 平滑肌 胰蛋白酶 肌动蛋白类 抗体 单克隆 荧光免疫测定 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]