pcDNA_4-PDGF-B重组质粒的构建并转染体外培养兔骨髓间充质干细胞  

Construction of Recombinant Plasmid of pcDNA_4-PDGF-B and Transfecting It to Rabbit Bone Marrow Mesenchymal Stromal Cells in Vitro

作　　者：赵松[1] 祝新平 龙清云[1] 周军[1] 张一帆[1] 张晓龙[1] 

机构地区：[1]武汉大学中南医院放射科,湖北武汉430071 [2]湖北省孝感市第一人民医院放射科,湖北孝感432000

出　　处：《武汉大学学报（医学版）》2013年第1期19-23,共5页Medical Journal of Wuhan University

基　　金：湖北省自然科学基金资助项目(编号:2010CDB06104)

摘　　要：目的:探讨自行构建pcDNA_4-PDGF-B重组质粒并转染兔骨髓间充质干细胞(BMSCs)的方法及可行性,并观察转染阳性细胞形态及生物特性。方法:构建包含人类血小板源性生长因子B(PDGF-B)重组质粒,将目的基因转染到兔BMSCs中,并通过形态观察、双酶切、聚合酶链反应、免疫组化来检测该目的基因在兔BMSCs中表达,通过G418抗生素筛选、MTT法绘制细胞生长曲线探讨PDGF-B基因对靶细胞生长影响。结果:pcDNA_4-PDGF-B重组质粒的构建合成及人PDFG-B基因成功转染BMSCs,并持续稳定地表述PDGF-B。结论:PDGF-B基因可以对骨髓间充干细胞进行转染,PDGF-B对靶细胞增殖、分裂有明显促进作用,为后期研究植入人PDGF-B转染BMSCs以缓解急性肝功能衰竭奠定了基础。Objective： To construct recombinant plasmid of pcDNA4-PDGF-B, and to investigate the ap- proach and possibility of transferring PDGF-B gene into rabbit bone marrow stromal cells （BM- SCs）. Methods： Recombinant plasmid harboring the human of PDGF-B gene was constructed and transfected into BMSCs, the results were identified by means of morphological observation, PCR, and immunohistochemistry. The changes in the biological characteristics of transfected BMSCs were also observed. BMSCs were screened by G418, and growth curve of the cells was analyzed to observe the influence of PBGF-B. Results： Stable over-expression of PDGF-B showed successful transfection to BMSCs and the construction of pcDNA4-PDGF-B. Conclusion： Stable expression of PDGF-B gene is acquired through transfecting BMSCs, and PDGF-B can improve the proliferation and mitosis of BMSCs. These results are helpful for the future treatment of acute liver failure by transplanting BMSCs with PDGF-B gene.

关 键 词：骨髓间充质干细胞 血小板源性生长因子-B 重组质粒构建 转染 

分 类 号：Q813[生物学—生物工程]