中国组织工程研究Issue(36)：5853-5858,6.DOI:10.3969/j.issn.2095-4344.2015.36.021
腺相关病毒介导β-神经生长因子基因转染内皮祖细胞
Transfection of adeno-associated virus encoding beta-nerve growth factor into endothelial progenitor cells
杨中雁 1高春正 1吴东进 1彭长亮1
作者信息
- 1. 山东大学第二医院，山东省济南市 250033
- 折叠
摘要
Abstract
BACKGROUND:Nowadays, gene therapy has become a new trend for disease therapy and brought promise for some refractory diseases. Its key is to choose proper cel s, genes and vectors. OBJECTIVE:To use recombinant adeno-associated virus mediatedβ-nerve growth factor (β-NGF) to transfect rat bone marrow-derived endothelial progenitor cel s in vitro, and to investigate the effect ofβ-NGF expression on the proliferation of endothelial progenitor cel s. METHODS:The endothelial progenitor cel s were isolated, cultured and identified from the bone marrow of rats. Empty vector or recombinant adenovirus-associated virus containingβ-NGF gene was transferred into endothelial progenitor cel s. We examined the transfection efficiency by fluorescence expression of green fluorescent protein. Expression ofβ-NGF protein was detected using ELISA, and its effect on the proliferation of endothelial progenitor cel s was determined using MTT method. RESULTS AND CONCLUSION:Rat endothelial progenitor cel s were isolated and cultured successful y in vitro and were identified positive by the function of cel s and immunofluorescence staining. The endothelial progenitor cel s were infected directly by the recombinant adenovirus-associated virus containingβ-NGF gene with an efficiency of 65.3%.β-NGF protein was detected in the culture supernatant of transfected endothelial progenitor cel s, which reached a high level at 10 days after gene transfection. Furthermore, there was noβ-NGF protein in the blank and empty vector groups. After transfection, the proliferative ability of endothelial progenitor cel s was increased, which was significantly higher than the blank and empty vector groups (P<0.05). But there was no difference between the latter two groups (P>0.05). These findings suggest that recombinant adenovirus-associated virus containingβ-NGF gene can be successful y transferred into rat bone marrow-derived endothelial progenitor cel s and promote the proliferation of endothelial progenitor cel s.关键词
干细胞/培养/神经生长因子/内皮祖细胞/基因转染/细胞培养/大鼠分类
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杨中雁,高春正,吴东进,彭长亮..腺相关病毒介导β-神经生长因子基因转染内皮祖细胞[J].中国组织工程研究,2015,(36):5853-5858,6.
