腺病毒载体携带标记基因EGFP转染人羊膜上皮细胞
Human amniotic epithelial cells transfected by enhanced green fluorescent protein gene mediated by adenovirus vector
摘要
Abstract
BACKGROUND:Human amniotic epithelial cells have some properties of stem cells, which can be induced to differentiate into corneal epithelial cells, but cannot be tracedin vitro. OBJECTIVE:To investigate the feasibility and infection efficiency of adenovirus vector carrying enhanced green fluorescent protein (EGFP) into the human amniotic epithelial cells. METHODS:The adenovirus vectors carrying EGFP was transferred into human amniotic epithelial cells culturedin vitro. After cultured and amplified, the morphology difference between transfected and non-transfected human amniotic epithelial cels was observed. The transfected human amniotic epithelial cells were observed under fluorescence microscope, and the cell cycle and the expression rate of EGFP in transfected human amniotic epithelial cells were detected by flow cytometry. RESULTS AND CONCLUSION:No obvious difference in the cell morphology was found between transfected human amniotic epithelial cells and normal human amniotic epithelial cells cultured in vitro. Flow cytometry analysis revealed that the EGFP positive rate was highest and reached up to 99.01% at 48 hours after transient transfection. The cell cycle of human amniotic epithelial cells transfected by the adenovirus vector was slowed a bit. To conclude, the adenovirus vector is a good medium of transfecting EGFP into human amniotic epithelial cells, and makes it more convenient to observe the further transformation of human amniotic epithelial celsin vitro.关键词
干细胞/培养/腺病毒/绿色荧光蛋白/人羊膜上皮细胞/原代培养/扩增培养/基因转染/转染效率/细胞周期/国家自然科学基金分类
医药卫生引用本文复制引用
金玲,刘小勇,徐玮,郝晓宁,牛静宜,王乙婷,曹端荣..腺病毒载体携带标记基因EGFP转染人羊膜上皮细胞[J].中国组织工程研究,2017,21(21):3382-3387,6.基金项目
国家自然科学基金项目(30872808,81100637) （30872808,81100637）
广东省医学科学技术研究基金项目(201512902216517) （201512902216517）
广东省科技计划项目(2015A020212026) （2015A020212026）
广东省中医药局科研项目(20162044) （20162044）
深圳市科技计划基金项目(JCYJ20140414105820176, CXZZ20140418182638764) the National Natural Science Foundation of China, No. 30872808, 81100637 （JCYJ20140414105820176, CXZZ20140418182638764）
the Medical Science and Technology Research Foundation of Guangdong Province, No. 201512902216517 （）
the Science and Technology Plan Project of Guangdong Province, No. 2015A020212026 （）
the Scientific Research Project of Guangdong Provincial Administration of Traditional Chinese Medicine, No. 20162044 （）
the Science and Technology Plan Project of Shenzhen City, No. JCYJ20140414105820176, CXZZ20140418182638764 （）
