中国病理生理杂志2009，Vol.25Issue(11)：2197-2203,7.
慢病毒载体介导2次转基因对人胚胎干细胞特性的影响
Effect of double gene transduction mediated by lentiviral vectors on characteristics of human embryonic stem cells
摘要
Abstract
AIM: To study the effect of double gene transduction mediated by lentiviral vectors on the characteristics of human embryonic stem cells (hESCs). METHODS: Using the backbone from inducibal dual and excisable transgene vector (iDuet101) , lentiviral vectors overexpressing cytotoxic T lymphocyte - associated molecule - 4 immuno-globulin (iDuet101 - CTLA4Ig) , indoleamine 2, 3 dioxygenase (iDuet101 - IDO) , and ubiquitin - C promoter - lueifer-ase - ires - puromycin ( ULIP) were constructed and packaged according to the standard recombinant techniques. Human embryonic stem cells (hESCs) were first transduced with iDuet101, iDuet101 - CTLA4Ig or iDuet101 - IDO, then, after the selection, were transduced again with ULIP. The expression and function of the exogenous genes were detected. Immu-nohistochemistry, RT - PCR and flow cytometry were applied for detection of embryoid bodies ( EB) formation in vitro and in vivo teratoma formation. RESULTS: Double - transduced hESCs showed typical shape of cell clones and positive staining of tumor rejection antigen -1 - 60 ( Tra -1 - 60 ) and octomer transcription factor - 4 ( OCT - 4 ). The formation of EB was observed, in which a - fetoprotein (AFP), paired box gene 6 ( Pax6) and Musashi 1 ( MSI1) were positively expressed. The cells formed teratomas, and the luciferase signals existed until 28 days after xeno - transplantation. CONCLUSION : Double transduction of non - transcriptional factors mediated by lentiviral vectors does not affect the cell growth rate and their differentiation ability.关键词
人胚胎干细胞/慢病毒载体/基因转染Key words
Human embryonic stem cells/ Lentiviral vector/ Gene transfection分类
医药卫生引用本文复制引用
蔡柳洪,周灿权,张滨,陶欣,林慧..慢病毒载体介导2次转基因对人胚胎干细胞特性的影响[J].中国病理生理杂志,2009,25(11):2197-2203,7.基金项目
863计划资助项目(No. 200601020A1022) （No. 200601020A1022）
