不同细胞生长因子组合对内皮祖细胞培养过程的影响

目的 观察不同细胞生长因子组合对体外培养过程中内皮祖细胞（EPC）数量和功能的影响。方法采用密度梯度离心法分离人脐血单个核细胞，接种至包被有人纤维连接蛋白的培养皿，按照处理方式不同分为4组：对照组（不添加细胞因子）、VEGF组加入血管内皮生长因子（VEGF）】、VEGF＋ECGS组加入VEGF及内皮细胞生长添加剂（ECGS）]、多因子组（加入多种细胞生长因子），6d后倒置显微镜下观察各组细胞集落形成情况，测定黏附能力及收获的EPC数量。结果VEGF组及ECGS＋VEGF组细胞集落形成能力（11.53±1.72、1310±2．09）均显著强于对照组（587±0．55）（均P〈0.01】；ECGS＋VEGF组细胞黏附能力（55．53±18.32）显著强于VEGF组（24.92±4．31）（均P〈0．01）；多因子组细胞集落形成及粘附能力（33．83±5．10、65.22±10、98）均明显强于其余各组（P〈0．05或0．01）。VEGF组及VEGF＋ECGS组收获的EPC数量分别为对照组的185、284倍，均显著增加（均P〈0.01）；多因子组EPC数量虽多于VEGF＋ECGS组，但差异并无统计学意义（P〉0．05）。结论EPC体外培养时，添加VEGF及ECGS是一种经济有效的方法，可显著改善EPC的集落形成及黏附能力，增加EPC的收获数量。

Different cell growth factors combination screening in EPC cell culture process

Objective To investigate the effects of various combination of cells growth factors on number and activity of endothelial progenitor cells （EPCs）.Methods Mononuclear fraction of human umbilical cord blood was obtained by density gradient centrifugation, and plated on culture dishes coated with fibronectin. Cells were divided into four groups： group C （control）, group V （supplemented with VEGF）, group VE （supplemented with VEGF and ECGS）, group M （supplemented with VEGF, FGF-β, IGF-1, G-CSF, EGF and flk3-Ligand ）. After six days of culture, cell clusters were observed with an inverted microscope, the adhesion ability were detected and the number of obtained EPCs was counted. Results Compared with group C, the cluster numbers of group V and group VE, were significantly increased（ p〈0.01 ）. Numbers of adhesive EPCs of group VE was significantly increased than that of group V. Compared with other groups, cluster numbers of adhesive EPCs of group M were highest. Compared with control group, the number of obtained EPCs of group V and group VE were increased by 1.85 and 2.84 fold respectively （P 〈0.01 ）. Conclusion VEGF and ECGS can effectively expand EPCs growth and enhance its activities in vitro.