碱性成纤维细胞生长因子基因转染优化脐带间充质干细胞的培养  

Effects of basic fibroblast growth factor gene transfection on the proliferation of umbilical cord mesenchymal stem cells

作　　者：郑颖娟[1] 田建昌[2] 

机构地区：[1]解放军第三七一中心医院肿瘤血液科,河南省新乡市453000 [2]解放军第三七一中心医院心胸外科,河南省新乡市453000

出　　处：《中国组织工程研究》2013年第45期7885-7890,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:目前多采取重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染的方法,将外源性碱性成纤维细胞生长因子基因转入脐带间充质干细胞内并持续表达,调控细胞增殖及定向分化,取得高效持久的治疗作用。目的:了解采用重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染对体外培养的脐带间充质干细胞增殖及细胞周期的影响。方法:体外培养脐带间充质干细胞,经重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染,分为对照组、空载病毒组、碱性成纤维细胞生长因子转染组。用RT-PCR,Western blot检测脐带间充质干细胞在转染前后碱性成纤维细胞生长因子基因和蛋白的表达。应用细胞生长曲线、CCK-8观察细胞生长的优化作用,采用流式细胞术测定细胞周期分布的变化。结果与结论:转染碱性成纤维细胞生长因子后,转染组与对照组、空载病毒组相比碱性成纤维细胞生长因子基因和蛋白水平均有表达,细胞的生长速度明显增快,细胞周期G0/G1期减少,S期细胞数增多,各组间差异有显著性意义(P<0.05)。说明,通过重组腺相关病毒作为载体介导碱性成纤维细胞生长因子基因转染能促进体外培养的脐带间充质干细胞增殖,对其培养有优化作用。BACKGROUND： At present, exogenous basic fibroblast growth factor gene can be transfected into umbilical cord mesenchymal stem cells via a recombinant adeno-associated virus vector and exhibit sustained expression in transfected cells. This method can regulate cell proliferation and directed differentiation to obtain efficient long-lasting therapeutic effects. OBJECTIVE： To investigate the effects of basic fibroblast growth factor gene transfection via a recombinant adeno-associated virus vector on the proliferation and cell cycle of human umbilical cord mesenchymal stem cells cultured in vitro. METHODS： Human umbilical cord mesenchymal stem cells were cultured by the suspension culture in vitro, and were trensfected by recombinant adeno-associated virus-mediated basic fibroblast growth factor gene. Cultured cells were divided into three groups： control group, basic fibroblast growth factor group, and recombinant adeno-associated virus group. Reverse trenscription-PCR and western blot were used to assess the knockdown efficiency. Cellular proliferation was determined by cell growth curve and Cell Counting Kit-8 assay. The cell cycle was analyzed by flow cytometry. RESULTS AND CONCLUSION： Compared with the other two groups, the expression of basic fibroblast growth factor mRNA and protein increased significantly, the cell growth speed was also significantly increased, the cell cycle of G0/G1 phase was decreased and cell number in S phase was increased in the basic fibroblast growth factor group after transfection. These findings suggest that the recombinant adeno-associated virus-mediated basic fibroblast growth factor gene can promote the proliferation of umbilical cord mesenchymal stem cells proliferation cultured in vitro, end also can optimize the cell culture.

关 键 词：干细胞 脐带脐血干细胞 脐带 间充质干细胞 碱性成纤维细胞生长因子 基因转染 腺病毒 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]