经转染人骨形态发生蛋白7基因后骨髓间充质干细胞mRNA的表达  

Expression of human bone morphogenetic protein7mRNA after the gene transfection in rabbit bone marrow-derived mesenchymal stem cells

作　　者：金丹[1] 曾位森[2] 裴国献[1] 齐凤菊[1] 魏宽海[1] 王珂[1] 陈滨[1] 

机构地区：[1]第一军医大学南方医院创伤骨科,广东广州510515 [2]第一军医大学细胞生物学教研室,广东广州510515

出　　处：《第一军医大学学报》2002年第3期222-225,共4页Journal of First Military Medical University

基　　金：国家重点基础研究发展规划973项目(1999054309)

摘　　要：目的构建人骨形态发生蛋白7(humanbonemorphogeneticprotein7,hBMP-7)逆转录病毒载体并检测经逆转录病毒介导基因转染的兔骨髓间充质干细胞(bonemarrow-derivedmesenchymalstemcell,MSCs)mRNA的表达。方法构建hBMP-7逆转录病毒载体后利用脂质体介导的基因转移技术转染包装细胞PT67,制备含目的基因的重组逆转录病毒液并感染兔骨髓间充质干细胞,采用原位杂交、RT-PCR检测hBMP-7mRNA在MSCs中的表达。结果成功构建了hBMP-7逆转录病毒载体,经hBMP-7基因转染的MSCs可表达外源性BMP-7mRNA。结论采用逆转录病毒介导的方法可以将hBMP-7转染至MSCs中并表达外源性基因。Objective To detecttheexpressionof humanbonemorphogeneticprotein7(hBMP-7)mRNAin rabbitbone marrow-derivedmesenchymalstemcellswithhBMP-7genetransfectionmediatedby retroviralvector.Methods Retroviral vectorfor hBMP-7genewas constructedthatwas transferredintothepackagingcellPT67mediatedby liposome.hBMP-7gene-positivecellcloneswereselectedwithG418(600ng/ml)andamplifiedto obtaintheretroviralsupernatantcontainingthe targetgenesthatweresubsequentlyusedto transfectrabbitbonemarrow-derivedmesenchymalstemcells(MSCs).HBMP-7mRNAexpressionin the MSCs was examinedby way of in situ hybridizationand reversetranscriptase-polymerasechain reaction.Results hBMP-7retroviralvectorwassuccessfullyconstructedandtransferredintoPT67cellswithresistanceto G418,andaftertransfectionwiththerecombinantretrovirus,transcriptionandexpressionof hBMP-7mRNAweredetectedin MSCs.Conclusion RabbitbonemarrowstemcellstransfectedwithhBMPgenevia retroviralvectorcan secretethe correspondent protein,indicatingthepossibilityof enhancingtheosteogeniccapacityof MSCsinthestudyof bonetissueengineering.

关 键 词：骨形态发生蛋白 骨髓间充质干细胞 基因表达 组织工程 mRNA HBMP-7 基因转染 

分 类 号：R680.2[医药卫生—骨科学]