摘要
目的:建立一个简便、廉价和稳定的小鼠淋巴管内皮细胞(LEC)培养体系.方法:应用不完全弗氏佐剂诱导小鼠腹腔淋巴管瘤形成,消化法分离获得LEC,置于自制的鼠尾胶包被的培养瓶(板)中培养.以免疫荧光化学法检测LEC特异性标记物VEGFR-3和LYVE-1的表达,以3H-TdR掺入率测定和细胞倍增时间检测LEC的增殖活力,以淋巴管形成试验判定LEC能否形成淋巴管样结构.结果:不完全弗氏佐剂能诱导小鼠腹腔淋巴管瘤形成,所获LEC活细胞数大于98%.免疫荧光化学检测表明,LEC表达VEGFR-3和LYVE-1.在鼠尾胶包被的培养瓶(板)中,LEC生长状况良好,3H-TdR掺入率明显增加,倍增时间为(42.7±3.2)h;在鼠尾胶凝胶中,LEC能形成淋巴管样结构.结论:鼠尾胶为贴黏剂的体系是一种简便、廉价和稳定的小鼠LEC培养体系.
AIM: To establish a convenient, cheap and stable culture system for mouse lymphatic endothelial cells (LEC). METHODS: Mouse lymphangiomas in abdominal cavity were induced by incomplete Freund's adjuvant, then disrupted and digested to obtain LEC, which were cultured in the flask or plate previously coated with rat-tail collagen. Expressions of LEC specific markers, VEGFR-3 and LYVE-1, were identified by immunofluorescence. The proliferation activity of LEC was evaluated by ^3H-TdR incorporation efficiency and cell doubling time. The capability of LEC to form lymphatic vessel-like structures was assessed by the in vitro lymphatic vessel formation assay. RESULTS: Mouse lymphangiomas in abdominal cavity were induced successfully by incomplete Freund's adjuvant, and more than 98% living LEC were harvested. The expressions of VEGFR- 3 and LYVE-1 were positive in LEC. In the rat-tail collagen coated flask or plate, LEC grew well, with 3 H-TdR incorporation efficiency increasing obviously and cell doubling time being (42.7 ± 3.2 ) h. In the gel formed by rat-tail collagen, LEC could form lymphatic vessel-like structures. CONCLUSION: Mouse LEC culture system using rat-tail collagen as adhesive is a convenient, cheap and stable culture system.
出处
《第四军医大学学报》
北大核心
2007年第5期390-393,共4页
Journal of the Fourth Military Medical University
基金
国家自然科学基金(30371586)
关键词
鼠尾胶
贴黏剂
内皮
淋巴管
细胞培养
rat-tail collagen
adhesive
endothelium, lymphatic
cell culture