CAJ | 학술논문

目的探讨干细胞因子(SCF)对脐静脉内皮细胞(HUVEC)增殖、迁移、管状形成能力的影响,以及对CD133+细胞的趋化效应.方法应用MTT及CCK-8增殖分析法检测HUVEC在不同细胞因子[空白试剂、SCF、血管内皮生长因子(VEGF)、抗人SCF、人IgG]条件下增殖能力的差异性;采用细胞划痕法与Matrigel体外三维成型法分别检测内皮细胞的增殖、迁移和管状形成能力;并应用Transwell技术检测不同细胞因子诱导的CD133+细胞体外趋化效应.结果 MTT及CCK-8增殖分析结果显示SCF无HUVEC增殖刺激活性;SCF可显著提升HUVEC迁移能力;SCF呈剂量依赖性增强HUVEC 管状形成能力,在适宜浓度SCF(100 ng/ml)作用下,HUVEC完整小管形成数量[(30.0 ±3.4)/105HUVEC]显著高于空白试剂组[(5.0±2.6)/105HUVEC,P<0.01];SCF可高效诱导CDl33+细胞体外趋化,SCF组[(118.0±6.5)/104CD133+细胞]Transwell小室跨膜迁移细胞数显著高于空白试剂组[(47.0±4.7)/104CDl33+细胞,P<0.01].结论 SCF可显著增强HUVEC的迁移及管状形成能力,并有效诱导CD133+细胞体外趋化,提示SCF/c-kit信号转导在内皮细胞及其祖细胞的血管新生与血管发生过程中可能发挥重要作用.
목적 탐토간세포인자(SCF)대제정맥내피세포(HUVEC)증식、천이、관상형성능력적영향,이급대CD133+세포적추화효응.방법 응용MTT급CCK-8증식분석법검측HUVEC재불동세포인자[공백시제、SCF、혈관내피생장인자(VEGF)、항인SCF、인IgG]조건하증식능력적차이성;채용세포화흔법여Matrigel체외삼유성형법분별검측내피세포적증식、천이화관상형성능력;병응용Transwell기술검측불동세포인자유도적CD133+세포체외추화효응.결과 MTT급CCK-8증식분석결과현시SCF무HUVEC증식자격활성;SCF가현저제승HUVEC천이능력;SCF정제량의뢰성증강HUVEC 관상형성능력,재괄의농도SCF(100 ng/ml)작용하,HUVEC완정소관형성수량[(30.0 ±3.4)/105HUVEC]현저고우공백시제조[(5.0±2.6)/105HUVEC,P<0.01];SCF가고효유도CDl33+세포체외추화,SCF조[(118.0±6.5)/104CD133+세포]Transwell소실과막천이세포수현저고우공백시제조[(47.0±4.7)/104CDl33+세포,P<0.01].결론 SCF가현저증강HUVEC적천이급관상형성능력,병유효유도CD133+세포체외추화,제시SCF/c-kit신호전도재내피세포급기조세포적혈관신생여혈관발생과정중가능발휘중요작용.
Objective 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. Methods 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-diamensional 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. Results 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)/105 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)/105HUVEC,P <0.01]. SCF also significantly induced a chemotactic response of CD 133+ cells, the transmembrane migration cell number into Transwell lower chamber was significantly higher in SCF group [(118.0 ±6.5)/104 CD133+ cells] than in blank control group [(47. 0 ±4. 7)/104 CD133 + cells,P <0.01]. Conclusions 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.