中文摘要：

背景：人羊膜上皮细胞具有干细胞的某些性能,可诱导分化为角膜样上皮细胞,但在体外培养时无法示踪,不能有效地监测其细胞转归。目的：探讨利用腺病毒载体将标记基因EGFP转染入人羊膜上皮细胞的可行性及感染效率。方法：pD C316-mC MV-EGFP腺病毒包装后,腺病毒载体携带标记基因EGFP转染体外培养的人羊膜上皮细胞,并扩增培养,观察转染后人羊膜上皮细胞与未转染人羊膜上皮细胞的形态差异,并在荧光显微镜下观察转染基因的表达,用流式细胞仪检测转染细胞的细胞周期及EGFP阳性表达率。结果与结论：（1）多数转染细胞筛选出的抗性克隆以及培养的克隆细胞与正常体外培养的人羊膜上皮细胞在形态上无显著差异;（2）瞬时转染后48 h的人羊膜上皮细胞中增强型绿色荧光蛋白阳性表达率最高达99.01%;（3）转染后的人羊膜上皮细胞的细胞周期有所减慢,说明利用腺病毒载体能将标记基因EGFP转染入人羊膜上皮细胞,可更直观地观察到人羊膜上皮细胞在体外的进一步转归。

英文摘要：

BACKGROUND： Human amniotic epithelial cells have some properties of stem cells, which can be induced to differentiate into corneal epithelial cells, but cannot be traced in 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 cultured in vitro. After cultured and amplified, the morphology difference between transfected and non-transfected human amniotic epithelial cells 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 cells in vitro.