The Scf On Huvecs Proliferation, Migration, Tube Formation Of Functional Impact And The Chemotactic Effect On Epcs

Objectives:SCF,a c-Kit ligand,is also called MGF or SLF.And c-Kit,as SCF receptor, is expressed in mature endothelial cells and endothelial progenitor cells.In order to find whether SCF possibly plays a important role in angiogenesis of vascular endothelial cells and in vasculogenesis of EPCs,we explore the effects of SCF on the proliferation,migration,capillary tube formation of HUVECs and the chemotaxis of EPCs.Methods:Endothelial cells were isolated from human umbilical vein by the digestive fluid,0.1%collagenaseⅠ;And every experiment was divided into 5 groups:blank control,SCF,VEGF,anti-SCF and IgG;Then,proliferation,migration and capillary tube formation of HUVECs were detected by MTT,CCK-8 analysis,and wound scratch assay, three-diamensional in vitro Matrigel assay,respectively.Furthermore,we sorted CD133~+ cells(EPCs) from human mononuclear cells of umbilical cord blood by flow cytometry, and investigated the chemotaxis of EPCs chemotaxis by transwell chemotaxis assays.Results:①Being digested by 0.1%Ⅰcollagenase for 15 minutes was the best choice to isolate ehdothelial cells from human umbilical vein;②SCF wasn't able to improve the proliferative ability of HUVECs,compared with blank control group,the proliferation rates in SCF(100ng/ml) group were 99±4.4%(MTT) 101±2.3%(CCK-8), (P＞0.05);③SCF had a significant role in enhancing the migration capacity of HUVECs;④SCF dose-dependently drove capillary tube formation of HUVECs,and the optimal concentration of SCF was 100ng/ml,the number of intact tubules formed on matrigel with an assay medium containing 100ng/mlSCF was 30±3.4,remarkably higher than the number of blank control group(5±2.6),(P＜0.05).⑤EPCs could be successfully obtained by sorting the cells which were labeled with fluorescent antibody CD133 from human mononuclear cells of umbilical cord blood by flow cytometry.⑥SCF markedly induced a chemotactic response of EPCs,compared to the results obtained from blank control group(47±4.7),the number of the cells of transmembrane migration in SCF group was up to 118±6.5(P＜0.05). Conclusions:Our study indicated that SCF promoted the migration,capillary tube formation of HUVECs,and also induced a chemotactic response of EPCs.This study also confirmed that SCF/c-Kit signaling possibly played a critical role in regulating angiogenesis of vascular endothelial cells and vasculogenesis of EPCs.