pcDNA_(3.1)-成骨生长肽真核表达载体在骨髓基质干细胞中的表达(英文)  

pcDNA_(3.1)-osteogenic growth polypeptide eukaryotic expression vector in bone marrow mesenchymal stem cells

作　　者：安刚[1] 吕松岑[1] 郭亚山[1] 薛震[1] 邓秋奎[1] 

机构地区：[1]哈尔滨医科大学附属第二医院骨科,黑龙江省哈尔滨市150086

出　　处：《中国组织工程研究与临床康复》2011年第36期6696-6700,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：the Returned Students Fund of Heilongjiang Province,No. LC04C03;the Youth Scientific Research Fund of Harbin City,Training Reserve Leader of Subject Fund,the Returned Students Fund Project,No. 2005AFLXJ016~~

摘　　要：背景:成骨生长肽具有明显的促进成骨细胞增殖、分化、成熟的作用。目的:观察成骨生长肽基因转染兔骨髓基质干细胞后的表达及表达产物对骨髓基质干细胞向成骨细胞分化的影响。方法:构建重组真核表达载体pcDNA3.1-成骨生长肽,并在脂质体介导下,将其导入兔骨髓基质干细胞。通过G418筛选获得阳性克隆。RT-PCR方法检测成骨生长肽基因在骨髓基质干细胞内的表达,并观察转染pcDNA3.1-成骨生长肽后骨髓基质干细胞向成骨细胞分化的情况。结果与结论:实验成功构建了真核表达载体pcDNA3.1-成骨生长肽,转染pcDNA3.1-成骨生长肽的骨髓基质干细胞可见成骨生长肽mRNA的表达,同时羟脯氨酸的分泌水平增加,碱性磷酸酶活性增高。证实经pcDNA3.1-成骨生长肽转染的兔骨髓基质干细胞不仅可以表达成骨生长肽,而且具有向成骨细胞分化的特性。BACKGROUND：Osteogenic growth polypeptide（OGP） had clear effect on promoting osteoblast proliferation,differentiation and mature.OBJECTIVE：To explore the expression of OGP gene,which was transfected into rabbit bone marrow mesenchymal stem cells（BMSCs） and to evaluate the effects of OGP on differentiation of rabbit BMSCs.METHODS：pcDNA3.1-OGP was constructed using gene cloning and recombination techniques.Rabbit BMSCs were transfected with pcDNA3.1-OGP mediated by lipofectamine 2000.The transfection positive cell clones were selected with G418.The expression of OGP gene was detected using reverse transcription-polymerase chain reaction analysis on an mRNA level.Differentiation of pcDNA3.1-OGP transfected BMSCs into osteoblast lineage was observed.RESULTS AND CONCLUSION：The pcDNA3.1-OGP plasmid was constructed successful and OGP expression was detected in rabbit BMSCs.Hydroxyproline content was increased,and alkaline phosphatase activity was also increased.These indicate that pcDNA3.1-OGP transfected BMSCs expressed OGP,and could differentiate into osteoblast lineage.

关 键 词：成骨生长肽 pcDNA3.1载体 骨髓基质干细胞 基因转染 表达 

分 类 号：R394.2[医药卫生—医学遗传学]