Efficient Genetic Modification of Cynomolgus Monkey Embryonic Stem Cells with Lentiviral Vectors

Cell Transplantation

Article

Chang Liu¹  Weiqiang Li¹  Jie Qin¹  Jing Chen¹  Xinbing Yu¹  Rui Chen¹  Andy Peng Xiang²  Bruce T. Lahn³  Li Zhang⁴  Guifu Wu⁵  Yongshui Fu⁶ 

[1] Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China; The Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou, P.R. China;Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China; The Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou, P.R. China; Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China; State Key Laboratory of Ophthalmology, Sun Yat-sen University, Guangzhou, P.R. China;Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China; The Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou, P.R. China; Department of Human Genetics and Howard Hughes Medical Institute, University of Chicago, Chicago, IL, USA;Department of Human Genetics and Howard Hughes Medical Institute, University of Chicago, Chicago, IL, USA;Division of Cardiology, First Affiliated Hospital, Sun Yat-sen University, Key Laboratory on Assisted Circulation, Ministry of Health, Guangzhou, P.R. China;Guangzhou Blood Center, Guangzhou, P.R. China;

关键词: Primate embryonic stem cells;    Lentivirus;    Transduction;    Green fluorescent protein;   

DOI  :  10.3727/096368910X504469

 received in 2009-08-11, accepted in 2010-04-20,  发布年份 2010

来源: Sage Journals

【 摘 要 】

Embryonic stem (ES) cells have the ability to undergo indefinite self-renewal in vitro and give rise during development to derivatives of all three primary germ layers (ectoderm, endoderm, and mesoderm), which make them a highly prized reagent in cell and gene therapy. Efficient introduction of various genes of interest into primate ES cells has proven to be difficult. Here, we demonstrated that the self-inactivating HIV-1-based lentiviral vectors constructed by MultiSite gateway technology are efficient tools for the transduction of cynomolgus monkey (Macaca fasicularis) ES (cmES) cells. After antibiotic selection, all of the transduced cells can stably express the reporter gene (humanized Renilla GFP or dTomato) while maintaining their stem cell properties, including continuous expression of stem cell markers, alkaline phosphatase (AKP), OCT-4, SSEA-4, and TRA-1-60, formation of embryoid bodies in vitro and teratomas in vivo containing derivatives of three embryonic germ layers. This approach will provide a useful tool for both gene function studies and in vivo cell tracking of stem cells.

【 授权许可】

Unknown   
© 2010 Cognizant Comm. Corp.

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