内皮生长因子对低温冻存内皮生长晕细胞增殖及黏附能力的影响

The effect of vessel endothelial growth factor on the proliferation and adherent ability of cryopreserved endothelial outgrowth cells

目的:探讨不同浓度血管内皮生长因子(vessel endothelial growth factor,VEGF)对内皮生长晕细胞(endothelial outgrowth cell,EOC)低温冻存过程中细胞生物学功能的保护作用。方法:采用密度梯度离心法分离脐血中的单个核细胞,培养扩增EOC,免疫组化、荧光染色及流式细胞仪检测其内皮细胞特性。第2代细胞用含0、10、25、50ng/mL VEGF的冻存液冻存,24h后复苏,CCK-8法检测增殖能力;差速贴壁法检测黏附能力。结果:CD34+、KDR+、VE-Cadherin+细胞分别为(61.17±0.44)%、(34.79±6.31)%、(74.64±6.96)%。10ng/mL VEGF组与空白对照组相比,其增殖能力无统计学差异,而黏附能力得到一定程度的保护。25ng/mLVEGF和50ng/mL VEGF组细胞复苏后的细胞增殖能力与黏附能力均较空白对照组增强。结论:VEGF能够不同程度地保护EOC在低温冻存中的生物学功能。

Objective To investigate the effect of vessel endothelial growth factor（VEGF） on the proliferation and adherent abilities of endothelial outgrowth cell（EOC） after cryopreservation.Methods EOC were cryopreserved using culture medium with or without 50 ng/mL VEGF.After 24 h,cryopreserved EOC were resuscitated,and the biological abilities were analysed.Results The percentage of CD34＋,KDR＋,VE-Cadherin＋ cell was（61.17 ± 0.44）%,（34.79 ± 6.31）%,（74.64 ± 6.96）% respectively.There was no significant difference between 10 ng/mL group and control group in proliferation ability,while the adherent ability was improved by 10 ng/mL VEGF.Both proliferation and adherent ability were improved by 25 ng/mL and 50 ng/mL VEGF.Conclusion VEGF can enhance the proliferation and adherent ability of EOC after cryopreservation.