含绿色荧光蛋白基因的子宫内膜异位症体外模型的建立

目的 由腺病毒介导绿色荧光蛋白基因转染子宫内膜细胞,构建子宫内膜异位症体外模型.方法 取5例正常子宫内膜组织,分离子宫内膜基质细胞和腺上皮细胞,进行形态学观察和免疫荧光鉴定.用携带有绿色荧光蛋白基因的腺病毒转染细胞,比较转染后12、18、36、42h细胞的绿色荧光蛋白表达阳性率和凋亡率的差异.结果 分离培养获得高纯度的子宫内膜腺上皮细胞(90%)和基质细胞(接近100%).两种细胞均表达绿色荧光;与腺上皮细胞比较,基质细胞转染率明显增高(F=8.362,P=0.020),凋亡率明显降低(F=46.119,P<0.001);转染对细胞的凋亡率无明显影响(F=4.208,P=0.057).结论 利用腺病毒介导的绿色荧光蛋白基因载体转染子宫内膜腺上皮细胞和基质细胞,可获得相对稳定的体外荧光子宫内膜异位症模型.

Establishment of a model of endometriosis in vitro with endometrial cells expressing green fluorescent protein

Objective To reproduce a model of endometriosis with endometrial cells expressing green fluorescent protein (GFP) in vitro induced by adenovirus. Methods Five batches of normal endometrial samples were isolated and cultured, morphologically observed and identied with immunofluorescence method. The isolated endometrial cells were then transfected with green fluorescent protein (GFP) gene-associated adenovirus. GFP positive rate and apoptosis rate were determined by flow cytometry, and they were compared 12, 18, 36, and 42 hours after transfection. Results The endometrial glandular cells and stromal cells were successfully isolated in high purity (90% in glandular cells and approximately 100% in stromal cells). Green fluorescence protein was expressed in the both types of infected cells. Compared with endometrial glandular cells, GFP positive rate in stromal cells was higher (F=8. 362, P=0.020) and apoptosis rate was lower (F=46.119, P<0.001); transfection factor threw no significant influence on the apoptosis of the cells (F=4. 208, P=0.057). Conclusion A stable in vitro fluorescent model of endometriosis can be reproduced by transf ecting endometrial glandular and stromal cells with adenovirus-encoded GFP gene.