發(fā)布時間：2018-11-15 16:39

【摘要】：目的研究過表達(dá)血管內(nèi)皮生長因子(VEGF)對人脂肪干細(xì)胞(hADSC)的細(xì)胞表型、增殖能力和多向分化潛能的影響。方法利用脂質(zhì)體法將pIRES2-EGFP-VEGF質(zhì)粒轉(zhuǎn)染第2代hADSC,對照組為脂質(zhì)體空載體。免疫熒光染色驗證轉(zhuǎn)染成功,流式細(xì)胞術(shù)檢測兩組細(xì)胞表型的差異,MTT法檢測細(xì)胞增殖能力,誘導(dǎo)其成脂和成骨分化并進(jìn)行油紅O和茜素紅染色。結(jié)果轉(zhuǎn)染組可見EGFP和VEGF的表達(dá),對照組無EGFP表達(dá)。兩組細(xì)胞均高表達(dá)CD29、CD44、CD90,低表達(dá)CD31、CD45。轉(zhuǎn)染組hADSC的增殖能力顯著強(qiáng)于對照組,且誘導(dǎo)分化后的脂滴和鈣結(jié)節(jié)的數(shù)量和面積均顯著高于對照組。結(jié)論過表達(dá)VEGF可增強(qiáng)hADSC的增殖能力,促進(jìn)其成脂和成骨分化,但并未顯著改變細(xì)胞表型。
[Abstract]:Objective to study the effects of overexpression of vascular endothelial growth factor (VEGF) on the phenotype, proliferative ability and multipotential differentiation of human adipose stem cells (hADSC). Methods pIRES2-EGFP-VEGF plasmid was transfected into the second generation hADSC, control group by liposome method. The transfection was successfully verified by immunofluorescence staining. The phenotypic differences between the two groups were detected by flow cytometry. The proliferative ability of the cells was detected by MTT method and the adipogenic and osteogenic differentiation was induced and stained with oil red O and alizarin red. Results the expression of EGFP and VEGF was found in the transfection group, but no EGFP expression was found in the control group. High expression of CD29,CD44,CD90, and low expression of CD31,CD45. in both groups The proliferative 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 induced differentiation were significantly higher than those in control group. Conclusion overexpression of VEGF can enhance the proliferation of hADSC and promote its adipogenic and osteogenic differentiation, but it does not significantly change the phenotype of hADSC.
【作者單位】： 解放軍總醫(yī)院整形修復(fù)科;美國德克薩斯大學(xué)MD安德森腫瘤中心整形外科;美國貝勒醫(yī)學(xué)院醫(yī)學(xué)系腎臟學(xué)部;
【基金】：國家自然科學(xué)基金(81272123,81571864)
【分類號】：R329.2

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