摘要
目的研究犬外周血中CD34^＋／CD133^＋／VEGFR-3^＋淋巴管内皮祖细胞的生物学特征，探讨血管内皮生长因子-C（VEGF-C）／血管内皮生长因子受体3（VEGFR-3）信号途径在淋巴管内皮祖细胞向淋巴管内皮细胞分化中的作用。方法用密度梯度离心法从犬外周血分离单个核细胞，再用流式细胞仪从单个核细胞分选VEGFR-3^＋细胞。通过VEGF-C诱导使VEGFR-3^＋细胞向内皮细胞分化。在扫描电镜和透射电镜下观察细胞表面形态和超微结构，并在共聚焦激光扫描显微镜下观察特征性标志物的表达。结果VEGFR-3^＋细胞同时表达CD34和CD133。经流式细胞仪分析，外周血单个核细胞中CD34^＋／VEGFR-3^＋细胞的含量约为0.13％，VEGFR-3^＋／CD133^＋细胞的含量约为0.08％。CD34^＋／CD^33^＋／VEGFR-3^＋细胞的体积较大，直径约为15μm。经VEGF-C诱导后，细胞数目增多。细胞变为梭形，伸出板状伪足和许多丝状伪足。诱导后1周，细胞表达血管性血友病因子。诱导后2周，细胞表面出现小凹，细胞内可见Weibel-Palade小体。细胞表达淋巴管内皮细胞特异性标志物LYVE-1，CD133标志消失。结论在VEGF-C诱导作用下，外周血中的CD34^＋／CD133^＋／VEGFR-3^＋淋巴管内皮祖细胞能向淋巴管内皮细胞分化。VEGF-C／VEGFR-3信号途径在淋巴管内皮祖细胞的分化过程中起着重要作用。
Objective To investigate the biological properties of CD34^＋/CD133^＋/VEGFR-3^＋ lymphatic endothelial progenitor cells in peripheral blood and explore the effects of the VEGF-C/VEGFR-3 signaling pathway on differentiation of lymphatic endothelial progenitor cells to lymphatic endothelial cells. Methods Mononuclear cells were isolated from peripheral blood by density centrifugation with Percoll solution, and VEGFR-3^＋ cells were sorted from them with flow cytometry. Differentiation of VEGFR-3^＋ cells was indueed with VEGF-C. The morphology and ultrastructures of the cells were observed with scanning and transmiaaion electron microscopes. Expression of surface markers were examined with a confocal laser scanning microecope. Results VEGFR-3^＋ cells expressed CD34 and CD133 antigen. The percentage of CD34^＋/ VEGFR-3^＋ and VEGFR-3^＋/CD133^＋ cells were 0. 13% and 0. 08% of peripheral blood MNC respectively. The size of CD34^＋/CD133^＋/VEGFR-3^＋ cells was about 15 μm in the diameter. After induction with VEGF- C, they were increased. The cells were shuttle-like in shape and extended the lamellipodia and many filopodia. After 1 week induction with VEGF-C, they expressed coagulation factor Ⅷ related antigen, and at 2 week induction, they showed caveolae on the surface and Weibel-Palade body inside the cells. The specific lymphatic endothelial marker LYVE-1 was expressed on the cells, and no longer expressed CD133. Conclusions CD34^＋/CD133^＋/VEGFR-3^＋ lymphatic endothelial progenitor cells from peripheral blood may differentiate into lymphatic endothelial cells. The VEGF-C/VEGFR-3 signaling pathway has important effects on the differentiation of the lymphatic endothelial progenitor cells.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2007年第3期169-173,共5页
Chinese Journal of Hematology
基金
国家自然科学基金资助项目（30570948）
高等学校博士点专项科研基金资助项目（20030246036）
关键词
内皮
淋巴管
内皮生长因子
受体
血管内皮生长因子
犬
Endothelium, lymphatic
Endothelial growth factor
Receptors, vascular endothlial growth factor
Dog
