腺相关病毒介导绿色荧光蛋白基因体外转染人羊膜上皮细胞

Adeno-associated virus vector-medicated green fluorescent protein transfected human amniotic epithelial cells in vitro

背景:人羊膜上皮细胞易于获得,采集简单,是细胞移植和组织修复的理想种子细胞,目前国内外尚无关于标记、示踪体外培养的人羊膜上皮细胞的报道。目的:探讨腺相关病毒载体介导绿色荧光蛋白基因对体外培养的人羊膜上皮细胞的转染效果。方法:取人羊膜标本,以胰蛋白酶消化法分离培养人羊膜上皮细胞,采用含绿色荧光蛋白的腺相关病毒进行转染,检测其转染效率。结果与结论:人羊膜上皮细胞可成功地在体外进行原代和传代培养,经含绿色荧光蛋白的腺相关病毒颗粒转染后,可稳定高效表达绿色荧光蛋白,转染效率达58%。

BACKGROUND： Human amniotic epithelial cells （AECs） are easy to obtain and can function as ideal seed cells for cell transplantation and tissue repair. Currently, marking and tracing of human AECs remains rarely reported. OBJECTIVE： To explore the efficiency of adeno-associated virus （AAV） vector-medicated green fluorescent protein （GFP） on in vitro cultured human AECs transfection. METHODS： Human amnion samples were harvested and trypsinized to isolate human AECs. The AECs were transfected with AAV-GFP, and the transfection efficiency was detected. RESULTS AND CONCLUSION： Human AECs were successfully primary cultured and passaged in vitro. AAV-GFP-transfected AECs stably and highly expressed GFP, with a transfection efficiency of 58%.