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

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.