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A versatile tool for tracking the differentiation of human embryonic stem cells |
Weiqiang LI1,2, Jie QIN1,2, Xinyu LI3, Li ZHANG4, Chang LIU1,2, Fei CHEN1,2, Zifei WANG1,2, Lirong ZHANG5, Xiuming ZHANG1,2, Bruce T. LAHN1,2, Andy Peng XIANG1,2,6( ) |
1. Center for Stem Cell Biology and Tissue Engineering, Sun Yat-Sen University, No. 74 Zhongshan Road 2,Guangzhou 510080, China; 2. The Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou 510080, China; 3. Zhongshan Medical School, Sun Yat-sen University, Guangzhou 510080, China; 4. Department of Human Genetics and Howard Hughes Medical Institute, University of Chicago, Chicago, Illinois 60637, USA; 5. Department of Pathophysiology, Guangdong College of Pharmacy, Guangzhou 510006, China; 6. Cell Therapy Center, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China |
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Abstract The ability of human embryonic stem cells (hESCs) to undergo indefinite self-renewal in vitro and to produce lineages derived from all three embryonic germ layers both in vitro and in vivo makes such cells extremely valuable in both clinical and research settings. However, the generation of specialized cell lineages from a mixture of differentiated hESCs remains technically difficult. Tissue specific promoter-driven reporter genes are powerful tools for tracking cell types of interest in differentiated cell populations. Here, we describe the construction of modular lentivectors containing different tissue-specific promoters (Tα1 of α-tubulin; aP2 of adipocyte Protein 2; and AFP of alpha fetoprotein) driving expression of humanized Renilla green fluorescent protein (hrGFP). To this end, we used MultiSite gateway technology and employed the novel vectors to successfully monitor hESC differentiation. We present a versatile method permitting target cells to be traced. Our system will facilitate research in developmental biology, transplantation, and in vivo stem cell tracking.
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Keywords
human embryonic stem cells
lentivector
transduction
green fluorescent protein
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Corresponding Author(s):
XIANG Andy Peng,Email:xiangp@mail.sysu.edu.cn
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Issue Date: 01 October 2010
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