小鼠血管内皮细胞的培养、鉴定及巨噬细胞移动抑制因子对其促增殖作用的研究  

Cultivation and Identification of Mouse′ Aorta Vascular Endothelial Cells and Proliferation by Macrophage Migration Inhibitory Factor

作　　者：段朝霞[1] 陈魁君[1] 张洁元[1] 李兵仓[1] 王建民[1] 

机构地区：[1]第三军医大学附属大坪医院野战外科研究所第六研究室,创伤,烧伤,复合伤国家重点实验室,重庆400042

出　　处：《解放军医药杂志》2013年第1期10-13,共4页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金资助项目(81101422)

摘　　要：目的探讨小鼠血管内皮细胞的培养、鉴定方法及巨噬细胞移动抑制因子(MIF)对其增殖活性的调节作用。方法取小鼠主动脉,去除周围的脂肪和结缔组织,将动脉环剪成0.5 mm大小,放入基质胶处理的12孔培养板内,待细胞长满,用胰酶消化后,取单细胞悬液用抗CD146免疫磁珠纯化,再用CD31抗体鉴定细胞纯度,按104个细胞/孔加入96孔板培养,细胞70%～80%融合时用不同浓度MIF或MIF抑制剂处理细胞,48 h后用甲基噻唑基四唑(MTT)检测细胞增殖程度。结果经CD146磁珠纯化后细胞纯度为95%以上,MTT检测结果显示低剂量MIF能促进血管内皮细胞的增殖,并具有剂量效应关系,而高剂量MIF则与MIF抑制剂一样,抑制血管内皮细胞的增殖。结论成功建立了简单可行的小鼠血管内皮细胞的体外培养方法,并证实一定剂量范围内的MIF能促进血管内皮细胞的增殖,说明MIF在血管内皮细胞的多种病理生理反应中起重要作用。Objective To explore cultivation and identification of mouse＇ vascular endothelial cells （VEC） and ac- commodation on proliferation by macrophage migration inhibitory factor （MIF）. Methods Murine aorta VEC were got rid of periphery fat and connective tissue, cut arterial circle to 0.5 mm of circularity and put into 12 apertures of cuhural laminae treated with matrigel. The trypsin digestion method was used after topful cells. The single cell suspension was purified by anti- CD146 immunomagnetie beads. The cells purity was detected with anti-CD31 antibody. Ceils were processed by different con- centrations of MIF or MIF inhibitor when cells inoseulated at 70% -80% range, after cultivated by 104 cells/apertures added in- to 96 apertures of cultural laminae. The proliferation of the pure VECs populations was detected by methyl thiazolyl tetrazolium （MTT） after 48 h. Results The purity of VEC was 95% after purified by CD146 magnetic beads. Low close of MIF could pro- mote VECs proliferation significantly which proved by detected results of MTT and had dose-effect relationships, but high dose of MIF had the same role with MIF inhibitor which could restrain VECs proliferation. Conclusion The VEC is isolated suc- cessfully by CD146 magnetic beads with high purity. A certain dosage range of MIF can stimulate vascular endothelial cell pro- liferation. The MIF may play an important role in the vascular pathophysiological reaction.

关 键 词：巨噬细胞移动抑制因子 血管内皮细胞 细胞培养技术 剂量效应关系 小鼠 

分 类 号：R-322[医药卫生] R329.5