Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Chinese Journal of Neuromedicine》 2007-11
Add to Favorite Get Latest Update

Experimental study on transfection of adenovirus vector encoding green fluorescent protein into human bone marrow-derived mesenchymal stem cells

XU Gang XU Ru-xiang JIANG Xiao-dan ZHOU De-xiang YAO Peng-fei QIN Kun CAI Ying-qian ZOU Yu-xi QIN Ling-sha.Department of Neurosurgery;Neurosurgery Institute of Guangdong Province,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China  
Objective To explore the transfection of adenovirus encoding green fluorescent protein(Ad-GFP)vector into human bone marrow-derived mesenchymal stem cells(BMSCs),the dose-effect relationship between them and the impact on biological properties of BMSCs,and further investigate the feasibility of using Ad-GFP vector to construct gene-modifled BMSCs.Methods BMSCs were separated to be cultured in vitro.Flow cytometry was applied to detect the immune phenotype of cells;the adenovirus was obtained by packaging cell lines 293;BMSCs were transfected with Ad-GFP of various titers(1×10~3-1×10~(10)PFU/mL);cytometry was used to analyze the transfection efficiency;inverted microscope was applied to observe the morphological changes of cells;cell counting kit 8(CCK8)was applied to detect the activity of cell proliferation;β-mercaptoethanol was added after serum evacuation to induce directed differentiation of Ad-GFP-transfected BMSCs into neuron-like cells. Results CD34 and CD45,and CD29 and CD44,the surface markers of the third-to sixth-passage BMSCs were respectively negative and positive.When the virus titer was 1×10~7 PFU/mL,the transfection rate was 55% and when virus titers were 1×10~9 and 1×10~(10)PFU/mL,the transfection rate was 85%.In addition,pathological phenomena occurred to cells at the virus titer of 1×10~(10)PFU/mL.The fluorescent expression achieved its climax at 7 d and could still be seen at 28 d.BMSCs expressing Ad-GFP had three types of subpopulation structures under a fluorescence microscope;when the virus titer was≥1×10~6 PFU/mL,BMSCs proliferation was suppressed at the early stage,in a dose-dependent manner. BMSCs transfection with Ad-GFP could be differentiated into neuron-like cells with the induction ofβ-mercaptoethanol.Neuron-specific enolase(NSE)was positive.Conclusion Ad-GFP with suitable titers can infect BMSCs effectively,and has little impact on biological properties of cells and no impact on the differentiation-inducing function.BMSCs can serve as seed cells during the application of Ad-GFP vector system to the research of gene therapy.
【CateGory Index】: R329
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
Chinese Journal Full-text Database 10 Hits
1 WU Jing,LI Yu-feng,SONG Min-xun,HUANG Bing,MA Xiu-li,LI Feng(Institute of Poutlry Shandong Academy of Agricultural Sciences,Jinan 250023,China);Expression,purification and antiviral activity analysis of the chicken interferon alpha mature protein expressed in pichia pastoris[J];Shandong Agricultural Sciences;2006-01
2 CHENG Bang-zhao,YU Wei-yi,ZHOU Li-sha,XIE Jin-wen,CHENG Bao-yan (School of Animal Science and Technology,Anhui Agricultural University,Hefei 230036);Cloning and expressing of partial VP2 gene of IBDV[J];Journal of Anhui Agricultural University;2008-02
3 Bursal Disease Virus WantgChengming (Department of Animalltusbandary and vetrinarymedicine of Anhuiagrotechnical Teachers College) CaiBaoxiang ChengPuyan (Department of Animal Husabandary and Verterinary Medicine of Nanjing Agricultrual University);Purification and Identification of Infectious[J];;1992-02
4 Wang Chengming Chen Puyan Cai Baoxiang (Department of Animal Husbandry and Veterinary Medicine) (Nanjing University Agricultural);DEVELOPMENT OF NEUTRALIZATION MONOCLONAL ANTIBODIES AGAINST INFECTIOUS BURSAL DISEASE VIROUS[J];;1993-01
5 Wang Chengming (Animal husbandry and veterinary medicine) Cat Baoxiang and Cheng Pugan(Naming Agricultural Lniversity. 210095);Detecting IBDV by double-antibodied ELISA[J];JOURNAL OF ANHUI AGROTECHNICAL;1995-03
6 Xu Puting Wang Yimin Zhao ZijiangGong Zhangda You Zhiyong Institute of Virology, Chinese Academy of Preventive Medicine,Institute of Epidemiology of Yunnan Province.;ISOLATION AND IDENTIFICATION OFTWO ORBIVIRUSES IN YUNNANPROVINCE[J];Chinese Journal of Virology;1988-03
7 Xu Puting Wang Yimin Zhao Zijiang You Zhiyong (Institute of Virology, Chinese Academy of Preventive Medicine, Beijing);CHARACTERIZATION OF NEWLY ISOLATED ALPHAVIRUSES FROM YUNNAN[J];Chinese Journal of Virology;1988-04
8 Xu Puting Wang Yimin Zuo JianminLin Jiagwen Xu Pimu Institute of Virology, Chinese Academy of Preventive Medicine, Beijing Hospital of Dai Nationality Autonomous Prefecture of Xi Shuang Ban Na of Yunnan Province, Jinghong, Yunnan;NEW ORBIVIRUSES ISOLATED FROM PATIENTS WITH UNKNOWN FEVER AND ENCEPHALITIS IN YUNNAN PROVINCE[J];Chinese Journal of Virology;1990-01
9 Li Jingnain Du Nianxing Xu Weiyan( Nanjing Agricultural University, Nanjing);IMMUNE RESPONSE TO VIRAL HEMORRHAGIC DISEASE OF RABBIT[J];Chinese Journal of Virology;1990-02
10 Luo Tingrong Yu Kelun Wei Jiahuai(Guangxi Agricultural College);DETECTION OF NEWCASTLE DISEASE VIRUS ( NDV ) BY ABC-ELISA[J];Chinese Journal of Virology;1990-03
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved