血管内皮生长因子基因转染的人脂肪干细胞具有较强的增殖和分化能力

VEGF overespression promotes the proliferation and differentiation of human adipose-derived stem cells

目的研究过表达血管内皮生长因子(VEGF)对人脂肪干细胞(hADSC)的细胞表型、增殖能力和多向分化潜能的影响。方法利用脂质体法将pIRES2-EGFP-VEGF质粒转染第2代hADSC,对照组为脂质体空载体。免疫荧光染色验证转染成功,流式细胞术检测两组细胞表型的差异,MTT法检测细胞增殖能力,诱导其成脂和成骨分化并进行油红O和茜素红染色。结果转染组可见EGFP和VEGF的表达,对照组无EGFP表达。两组细胞均高表达CD29、CD44、CD90,低表达CD31、CD45。转染组hADSC的增殖能力显著强于对照组,且诱导分化后的脂滴和钙结节的数量和面积均显著高于对照组。结论过表达VEGF可增强hADSC的增殖能力,促进其成脂和成骨分化,但并未显著改变细胞表型。

Objective To investigate the effect of vascular endothelial growth factor（VEGF） overexpression on the phenotype,proliferation and multilineage differentiation ability of human adipose-derived stem cells（hADSCs）. Methods Passage-2 hADSCs were transfected with pIRES2-EGFP-VEGF plasmid using DOTAP liposomal technique, whereas hADSCs in control group were transfected in absence of plasmid. The success of transfection was confirmed by immunofluorescent cytochemistry. Phenotype differences between the two groups were detected using flow cytometry. Proliferation ability was assessed using MTT assay. Adipogenic and osteogenic differentiation were induced and confirmed using Oil Red O staining and alizarin red staining. Results The transfected hADSCs expressed EGFP and VEGF,whereas no EGFP was observed in the untransfected hADSCs. Both groups of hADSCs showed high expressions of CD29,CD44,CD90,and low expressions of CD31 and CD45. Compared with the untransfected hADSCs,the transfected hADSCs exhibited significantly increased proliferation ability as well as the number of lipid droplets, calcium nodules, and the stained area. Conclusion VEGF overexpression promotes proliferation and multilineage differentiation potential of hADSCs without evident phenotype changes.