三种裸鼠主动脉血管内皮细胞原代培养方法的比较  

Primary culture and identification of vascular endothelial cells of nude mice aorta: Comparison of three methods

作　　者：杨澜波 黄涛 史占军[2] 

机构地区：[1]150医院全军军事训练医学研究所骨科,河南洛阳471031 [2]南方医科大学南方医院关节骨病外科,广东广州510515

出　　处：《实用医药杂志》2012年第4期360-362,共3页Practical Journal of Medicine & Pharmacy

基　　金：国家自然科学基金(编号:30371450);广东省自然科学基金(编号:32948);广东省科技计划(编号:2008B030301348)

摘　　要：目的探索分离、培养裸鼠主动脉血管内皮细胞的有效方法。方法在无菌条件下从背部入路取得裸鼠的主动脉,分别通过贴壁消化法、动脉环法和植块法进行裸鼠主动脉内皮细胞的原代培养。经胰酶消化传代后,采用免疫细胞化学法鉴定血管内皮细胞标志分子Ⅷ因子在细胞中的表达情况。结果内壁贴壁消化法于培养后的5d后可见明显内皮细胞集落,约7～9 d后细胞汇合。其他两种方法细胞爬出出现较晚。内壁贴壁消化法和动脉环法内皮细胞标志物Ⅷ因子表达阳性率分别达98%和82%,而植块法只有58%。结论裸鼠主动脉内壁贴壁消化法和动脉环法操作性较强,可获取一定数量纯度较高的主动脉血管内皮细胞。Objective To explore an effective method of isolating and culturing the vascular endothelial cells of nude mice aorta.Methods The aorta was harvested under aseptic condition from the back of a nude mouse.Three different methods were used in primary culture of aortic endothelial cells in nude mice.After serial subcultivation,the VEC was identified by immunohistochemical method with anti-Ⅷ factor antibody.Results A small amount of cells were seen and adhered to the bottom of petri dishes 5 days after cultivation by digestion-adherent method,the VEC spreaded to confluent 7-9 days later.The other two methods were grew slower than digestion-adherent method.The VEC were identified as endothelial cells with 98 % expression of Ⅷ factor,which was regarded as the marker of endothelial cells,in rest two methods only 82% and 58%.Conclusion The digestion-adherent method and the explants method established in the present study are easy to handle,and can obtain a certain number of high purity VEC.

关 键 词：血管内皮细胞 裸鼠 原代培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]