Labeling of stable glioma stem cells with red fluorescent protein transfected by lentiviral vector
10.3760/cma.j.issn.1671-8925.2012.07.023
- VernacularTitle:红色荧光基因慢病毒转染人脑胶质瘤干细胞的实验研究
- Author:
Jin-Ming CHEN
1
;
You-Xin ZHOU
;
Ting-Feng WU
;
Xiong-Hui CHEN
;
San-Song CHEN
;
Zuo-Peng SU
;
Bin LI
;
Xue-Shun XIE
;
Gui-Lin CHEN
;
Yong-Xin WEI
;
Zi-Wei DU
Author Information
1. 赣南医学院第一附属医院
- Keywords:
Glioma stem cell;
Lentivirus;
Red fluorescent protein
- From:
Chinese Journal of Neuromedicine
2012;(7):737-740
- CountryChina
- Language:Chinese
-
Abstract:
[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 further basic research.
