TGF-β_1、bFGF双基因修饰骨髓间充质干细胞的实验研究  

Bone Marrow Mesenchymal Stem Cells Modified by TGF-β_1 and bFGF

作　　者：戴芳[1] 宋莉[1] 朱德星[2] 邹源 DAI Fang;SONG Li;ZHU De-xing;ZOU Yuan(Department of Stomatology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Oral Medicine,Stomatology Hospital of Guangzhou Medical University,Guangzhou 510140,China)

机构地区：[1]南昌大学第二附属医院口腔科,南昌330006 [2]广州医科大学附属口腔医院口腔内科,广州510140

出　　处：《南昌大学学报（医学版）》2018年第1期22-28,31,共8页Journal of Nanchang University：Medical Sciences

基　　金：江西省科技厅科技支撑项目(2010BSA15900)

摘　　要：目的以脂质体为载体来介导转化生长因子-β_1(TGF-β_1)、碱性成纤维细胞生长因子(bFGF)基因修饰骨髓间充质干细胞(BMSCs),观察两基因的表达情况。方法对SD大鼠的BMSCs采用贴壁法联合Percoll密度梯度离心法获得。使用PCR技术获得TGF-β_1基因、bFGF基因,将其基因在真核表达载体pcDNA3.1(+)上构建,再将两种真核表达载体通过脂质体转染至第3代BMSCs。采用RT-PCR、Western blot检测BMSCs中bFGF、TGF-β_1基因的表达水平。结果贴壁法联合Percoll密度梯度离心可以获得BMSCs。成功构建pcDNA3.1(+)-bFGF和pcDNA3.1(+)-TGF-β_1真核表达载体后,将其转染BMSCs,RT-PCR、Western blot均可检测到bFGF、TGF-β_1基因的表达。结论 bFGF和TGF-β_1可以通过脂质体转染方式修饰BMSCs,并获得高效表达。Objective To modify bone marrow mesenchymal stem cells(BMSCs)using liposome-mediated transforming growth factor-β1(TGF-β1)and basic fibroblast growth factor(bFGF)genes,and to observe the expression of the two genes.Methods The BMSCs were isolated from SD rats by adherence method combined with Percoll density gradient centrifugation.The TGF-β1 and bFGF genes were obtained by PCR to construct the eukaryotic expression vector pcDNA3.1(+).These two vectors were transfected into the third generation of BMSCs using the liposome.The expression of bFGF and TGF-β1 in BMSCs was detected by RT-PCR and Western blot.Results The BMSCs were obtained by adherence method combined with Percoll density gradient centrifugation.After transfection with the co nstructed pcDNA3.1(+)-bFGF and pcDNA3.1(+)-TGF-β1 eukaryotic expression vectors,the expression of bFGF gene and TGF-β1 was detected in BMSCs by both RT-PCR and Western blot.Conclusion The bFGF and TGF-β1 can be highly expressed in BMSCs modified by liposome transfection.

关 键 词：转化生长因子-Β1基因 碱性成纤维细胞生长因子基因 骨髓间充质干细胞 真核表达载体 脂质体转染 

分 类 号：R78[医药卫生—口腔医学] Q78[医药卫生—临床医学]