论文部分内容阅读
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 cell phenotype, proliferation and multidrug differentiation potential of human adipose-derived stem cells (hADSCs). Methods The pIRES2-EGFP-VEGF plasmid was transfected into the second generation of hADSC by liposome method. The control group was liposome empty vector. Immunofluorescence staining showed that the transfection was successful. The difference of cell phenotypes was detected by flow cytometry. The cell proliferation was detected by MTT assay. The adipogenic and osteogenic differentiation was induced by oil red O and alizarin red staining. Results The expression of EGFP and VEGF was observed in the transfected group, but not in the control group. Two groups of cells were highly expressed CD29, CD44, CD90, low expression of CD31, CD45. The proliferation ability of hADSC in transfection group was significantly stronger than that in control group, and the number and area of lipid droplets and calcium nodules after differentiation were significantly higher than those in control group. Conclusion Overexpression of VEGF can enhance hADSC proliferation and promote adipogenic and osteogenic differentiation, but did not significantly change the cell phenotype.