红色荧光基因慢病毒转染人脑胶质瘤干细胞的实验研究
陈金明;韦永新;杜子威;周幽心;吴庭枫;陈雄辉;陈三送;苏作鹏;李斌;谢学顺;陈桂林
【期刊名称】《中华神经医学杂志》 【年(卷),期】2012(000)007
【摘要】[Objective]To establish stable glioma stem cells with a high expression level ofred fluorescent protein (RFP) in vitro.[Methods]The glioma stem cells SU2 were transfected with lentivirus vector containing RFP gene;cell expression of RFP was observed by fluorescent microscopy;RFP-positive glioma stem cells were sorted out by fluorescence-activated cell sort.The transfection efficiency of SU2 cell before transfection and 20-passaged RFP-SU2 cells after transfection were assayed by flow cytometry.Dynamic viewing was performed to observe the cell division and cloning of RFP-SU2 single cell;RFP -positive cells were collected and immunostained with antibodies against CD133 and nestin.[Results] PFP-SU2 cells grew as levitated sphere with high RFP expression;the transfection efficiency of SU2 cell before transfection was only 1.5％,while that of20-passaged RFP-SU2 cells after transfection reached to 75％.RFP-SU2 single cell could proliferate into brain tumor stem cell spheres,having the abilities of self-renewing and clonal proliferation.Immunofluorescence showed positiveCDt 33 and nestin expressions in RFP-SU2 cells.[Conclusion] A SU2/RFP cell line marked by RFP is established;and it can serve as a promising tool for
红色荧光基因慢病毒转染人脑胶质瘤干细胞的实验研究
