绿色荧光蛋白和红色荧光蛋白共转染骨髓间充质干细胞、乳鼠心肌细胞、Eahy926细胞表达特征的共聚焦分析

Confocal microscopy analysis of expression characteristic of mesenchymal stem cells,cardiac myocyte and Eahy926 cells cotransfected by green fluorescent protein and red fluorescent protein

目的利用携带绿色荧光蛋白(GFP)、红色荧光蛋白(RFP)报告基因的慢病毒载体共转染,观察2种报告基因在人骨髓间充质干细胞(hMSC)、乳鼠心肌细胞(Myo)和内皮细胞株(Eahy926)中的整合表达特征。方法应用人类1型免疫缺陷病毒改造而构建慢病毒载体,将GFP、RFP以同时或先后方式共转染hMSC、乳鼠Myo和Eahy926细胞,激光共聚焦显微镜观察2种报告基因在不同细胞中的共表达特点。结果 3种细胞均能被GFP、RFP共转染,其中hMSC和乳鼠Myo均有竞争加随机转染方式,Eahy926只有随机转染方式;3种细胞转染GFP、RFP的效率有所不同,共表达2种报告基因的细胞表现为以核为中心的表达区域重叠。结论 hMSC、乳鼠Myo和Eahy926细胞均可共转染2种报告基因,但整合方式有所不同,这将对利用多个报告基因同时示踪细胞多种生命特征提供有力帮助。

Objective By using slow virus vector which take along green fluorescent protein（GFP）and red fluorescent protein（RFP）for cotransfection to observe the characters of the integrated expression of two reporter genes in human mesenchymal stem cells（hMSC）,cardiac myocytes（Myo） of neonate rat and endothelial cell line（Eahy926）.Methods The human immunodeficiency virus type 1 was constructed into lentivirus vectors.The hMSC,Myo and Eahy926 were transfected by GFP,RFP at the same time or in succession.Expression characteristic of two kinds of reporter gene in different cells were detected by confocal microscopy.Results Three kinds of cells can be cotransfected by GFP and RFP.There were competition and random transfection ways in hMSC and Myo cells,but there was only random transfection in Eahy926;There were different transfection efficiency of GFP and RFP and the coexpression of reporter gene had area overlapping signs with nuclear as the center.Conclusion All of hMSC,Myo and Eahy926 cells can cotransfection two kinds of report gene,but with different intergration ways,which will provide help in labellingtracing the cellular vital signs by multiple report gene.