人肾小球系膜细胞原代培养方法的改进

背景:肾小球系膜细胞原代培养成功率低,生存时间短,传代次数少.其中分离提取肾小球一直是培养高纯肾小球系膜细胞的最大瓶颈.目的:建立一种操作简单、成功率高、重复性好的人肾小球系膜细胞原代培养方法.方法:取自愿水囊引产的胎儿肾,采用肾皮质组织块法合优生选择法培养人肾小球系膜细胞.相差显微镜、透射电镜观察系膜细胞形态,免疫荧光技术鉴定细胞表型,激光共聚焦显微镜观察波形蛋白的表达.结果与结论:培养的肾小球系膜细胞呈梭形/不规则星形和细长形,胞质中各种细胞器丰富,细胞突起明显,胞膜上有很多微绒毛,细胞表达α-肌动蛋白、肌球蛋白、波形蛋白、结蛋白,而不表达细胞角蛋白、Ⅷ因子,激光共聚焦显微镜下发现系膜细胞波形蛋白表达阳性并具有特征性纤维束.说明培养的系膜细胞符合肾小球系膜细胞的生物学特征.肾皮质组织块法合优生选择法是一种简单、高效的培养人肾小球系膜细胞的方法.

A modified method for primary culture of human glomerular mesangial cells

BACKGROUND: Primary culture of glomerular mesangial cells was less achievement ratio, short survival time, and less passage times. In particular, extraction of renal glomerulus remains difficult for culturing highly pure mesangial cell. OBJECTIVE: To establish a more simple, high successful rate and good reproducibility method of human mesangial cells in primary culture. METHODS: Kidneys isolated from induction of labor with water bag voluntary were cut into pieces, and human mesangial cells were cultured w...