摘要
【目的】探索建立人子宫内膜异位症荧光体外与在体模型的具体方法。【方法】采用增强型绿色荧光蛋白腺病毒(Ad-eGFP)分别转染原代培养人子宫内膜腺上皮细胞和基质细胞(细胞转染注射法,即方法1)和子宫内膜组织块(组织转染注射法,即方法2),比较两种体外转染的差异;然后采用方法1将绿色荧光腺病毒转染后的腺上皮细胞和基质细胞注入裸鼠皮下形成荧光病灶,并且与方法2相比较,观察直至建模后25d,计算病灶荧光成模率与病灶荧光存在时间,并给予组织鉴定。【结果】方法1与2建模后5d两种方法病灶荧光阳性率分别达88.9%和22.2%,P=0.015;病灶体外荧光存在时间分别为(12±8)d与(7±4)d。病理组织学HE染色和免疫荧光共同鉴定显示病灶来源于人子宫内膜。【结论】应用Ad-eGFP转染后人子宫内膜腺上皮细胞和基质细胞混悬液注射可成功建立裸鼠皮下人子宫内膜异位症活体荧光观察模型,同时方法成模率高,体外荧光存在时间较长。
【Objective】 To establish a novel noninvasive fluorescent animal model for endometriosis in vitro and in vivo. 【Methods】 Adenovirus encoding enhancing green fluorescent protein （Ad-eGFP） was used to transfect endometrial glandular cells and stromal cells （cells transfection and injection, Method No.1）, and fragments （tissues transfection and injection, Method No. 2）. Transfection efficiencies were compared between the two methods in vitro. Then GFP transfected glandular cells and stromal cells suspension were injected into nude mice subcutaneously （Method No.1）, taking Method No.2 as a comparison. In vivo observation last for 25 days, and positive rates and duration times of fluorescent lesions were calculated. Histological examination was used to confirmed lesion formation. 【Results】 On the fifth day after injection, lesion positive rate of Method No.1 was 88.9%, which was statistically significantly higher than that of Method No.2 （22.2%）, P = 0.015 0.05. The fluorescent positive duration of Method No.1 and No.2 were 12 ± 8 days and 7 ± 4 days. The structures of lesions were all identified as human original endometrium by histological examination, including HE staining and immunofluorescency. 【Conclusion】 Noninvasive animal model of endometriosis can be built up by subcutaneously injection of Ad-EGFP transfected endometrial glandular cells and stromal cells suspension with higher positive rate and longer observation time
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2010年第2期298-301,共4页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金(30672222)
广东省自然科学基金(0400377
8151008901000119)
广东省科技计划项目(2006B50107001
2005B34201020
2007B080701011)
