Liu Rui, Tan Yu-Zhen, Wang Hai-Jie, Zhang Meihua
Sorting of lymphatic endothelial progenitor cells from canine peripheral blood and their differentiation induction towards endothelial cells
Zhonghua Xue Ye Xue Za Zhi. 2007 Mar;28(3):169-73.
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.
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 induced with VEGF-C. The morphology and ultrastructures of the cells were observed with scanning and transmission electron microscopes. Expression of surface markers were examined with a confocal laser scanning microscope.
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 microm in the diameter. After induction with VEGFC, 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 VII 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.
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.
研究外周血中CD34+/CD133 +/VEGFR-3 +淋巴管内皮祖细胞的生物学特性，探讨VEGF-C/VEGFR-3信号通路对淋巴管内皮祖细胞向淋巴管内皮细胞分化的影响。
采用Percoll溶液密度梯度离心法从外周血中分离单个核细胞，用流式细胞术从中分选VEGFR-3+细胞。用VEGF-C诱导VEGFR-3+细胞分化。用扫描电子显微镜和透射电子显微镜观察细胞的形态和超微结构。用共聚焦激光扫描显微镜检测表面标志物的表达。
VEGFR-3+细胞表达CD34和CD133抗原。CD34+/VEGFR-3+和VEGFR-3+/CD133+细胞在外周血单个核细胞中所占比例分别为0.13%和0.08%。CD34+/CD133+/VEGFR-3+细胞直径约为15微米。经VEGFC诱导后，细胞数量增加。细胞呈梭形，伸出片状伪足和许多丝状伪足。经VEGF-C诱导1周后，细胞表达凝血因子VII相关抗原，诱导2周后，细胞表面出现小凹，细胞内出现Weibel-Palade小体。细胞表达特异性淋巴管内皮标志物LYVE-1，不再表达CD133。
外周血中CD34+/CD133+/VEGFR-3+淋巴管内皮祖细胞可能分化为淋巴管内皮细胞。VEGF-C/VEGFR-3信号通路对淋巴管内皮祖细胞的分化具有重要作用。
