Yan Xin, Liu Bin, Lu Shi-hong, Ge Mei-li, Li Xing-xin, Zheng Yi-zhou
State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Tianjin 300020, China.
Zhonghua Xue Ye Xue Za Zhi. 2011 May;32(5):326-30.
To explore the effects of stem cell factor (SCF) on proliferation, transmigration, capillary tube formation of human umbilical vein endothelial cells (HUVEC) and on the chemotaxis of CD133(+) cells.
In the presence of blank control, SCF, vascular endothelial growth factor (VEGF), anti-human SCF (anti-SCF) or human IgG, the difference in proliferation capacity of HUVEC was analyzed by MTT and CCK-8 methods, and wound scratch assay and three-dimensional in vitro Matrigel assay were used for transmigration and capillary tube formation of HUVEC, respectively. In addition, the chemotaxis of CD133(+) cells sorted from human umbilical cord blood by flow cytometry was investigated by Transwell migration assay.
SCF didn't improve the proliferative capacity of HUVEC, but significantly enhanced the transmigration capacity, and increased capillary tube formation in a dose-dependent manner. The number of intact tubules [(30.0 ± 3.4)/10(5) HUVEC] formed by HUVECs in the presence of the optimal concentration of SCF (100 ng/ml) was remarkably higher than that in blank control group [(5.0 ± 2.6)/10(5) HUVEC, P < 0.01]. SCF also significantly induced a chemotactic response of CD133(+) cells, the transmembrane migration cell number into Transwell lower chamber was significantly higher in SCF group [(118.0 ± 6.5)/10(4) CD133(+) cells] than in blank control group [(47.0 ± 4.7)/10(4) CD133(+) cells, P < 0.01 ].
SCF significantly promotes the transmigration and capillary tube formation of HUVEC, and induces a chemotactic response of CD133(+) cells. SCF/c-kit signaling possibly plays a critical role in regulating angiogenesis of vascular endothelial cells and vasculogenesis of endothelial progenitor cells.
探讨干细胞因子(SCF)对人脐静脉内皮细胞(HUVEC)增殖、迁移、毛细血管管腔形成的影响以及对CD133(+)细胞趋化性的影响。
在空白对照、SCF、血管内皮生长因子(VEGF)、抗人SCF(抗-SCF)或人IgG存在的情况下,采用MTT法和CCK-8法分析HUVEC增殖能力的差异,分别采用划痕实验和体外三维基质胶实验检测HUVEC的迁移和毛细血管管腔形成能力。此外,采用Transwell迁移实验研究经流式细胞术从人脐带血中分选的CD133(+)细胞的趋化性。
SCF未提高HUVEC的增殖能力,但显著增强其迁移能力,并呈剂量依赖性增加毛细血管管腔形成。在最佳浓度SCF(100 ng/ml)存在的情况下,HUVEC形成的完整管腔数量[(30.0±3.4)/10(5) HUVEC]显著高于空白对照组[(5.0±2.6)/10(5) HUVEC,P<0.01]。SCF还显著诱导CD133(+)细胞的趋化反应,SCF组[(118.0±6.5)/10(4) CD133(+)细胞]跨膜迁移至Transwell下室的细胞数量显著高于空白对照组[(47.0±4.7)/10(4) CD133(+)细胞,P<0.01]。
SCF显著促进HUVEC的迁移和毛细血管管腔形成,并诱导CD133(+)细胞的趋化反应。SCF/c-kit信号通路可能在调节血管内皮细胞血管生成和内皮祖细胞血管发生中起关键作用。