GFP和RFP报道基因在人体不同细胞中转导率的比较研究  

Transduction Efficiency Comparison of GFP and RFP Reporter Gene in Human Cell Lines of hMSC,Eahy926 and A549

作　　者：罗毅[1] 辛毅[1] 张颖[1] 汪劲松[2] 王世强[3] 李景超[3] 高飞[4] 黄益民[1] 

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所,北京100029 [2]北京放射医学研究所,北京100850 [3]北京大学生命科学院,北京100048 [4]中国科学院过程工程研究所-生物工程国家重点实验室,北京100190

出　　处：《北京生物医学工程》2011年第1期20-25,共6页Beijing Biomedical Engineering

基　　金：北京市自然科学基金(5042006;7072012)资助

摘　　要：目的比较慢病毒载体介导的绿色荧光蛋白(GFP)和红色荧光蛋白(RFP)报道基因在人体不同细胞中的转导率,为再生医学和组织工程学研究中报道基因的选择提供依据。方法将构建的GFP和RFP慢病毒载体分别转染人骨髓间充质干细胞(hMSC)、人脐静脉内皮细胞株(Eahy926)和人肺泡上皮细胞株(A549)。4d后,用流式细胞仪检测GFP和RFP的转导率,并利用激光共聚焦显微镜观察GFP和RFP的表达特征。结果 GFP和RFP在hMSC、Eahy926和A549三种细胞间的转导率均有显著性差异(P(0.01);在hMSC或Eahy926或A549的同种细胞中GFP和RFP的转导率也有显著性差异(P(0.01);RFP在部分Eahy926和A549细胞局部高表达。结论慢病毒载体介导的GFP和RFP报道基因在人体不同细胞中的转导率明显不同。Objective By comparing the transduction efficiencies of GFP and RFP reporter genes in human cell lines hMSC, Eahy926 and A549, we provide the choice of reporter genes in the regeneration medicine and the organization engineering researches. Methods Human cell lines of hMSC, Eahy926 and A549 in vitro were respectively infected with lentiviral vectors encoding GFP and RFP reporter gene at MOI 100. After 4 days of transfection, the transduction efficiencies of GFP and RFP in each cell line were evaluated by flow cytometry. The expressive features of GFP and RFP reporters in cells were observed with confocal microscopy. Results We found that the transduction efficiencies of either GFP or RFP among three cell lines were very significantly difference （ P 〈 0. 01 ）. The transduction efficiencies of GFP and RFP among same cell line also was markedly varied and the difference was statistical significant （ P 〈 0.01 ）. The＂ RFP was concentrated expression at the subcellular localization in small population cells of Eahy926 and A549 strain.. Conclusion The transduetion efficieneies of GFP and RFP were evidently different among hMSC, Eahy926 and A549 cell lines.

关 键 词：绿色荧光蛋白(GFP) 红色荧光蛋白(RFP) 转导率 

分 类 号：R318[医药卫生—生物医学工程]