CAJ | 학술논문

[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25 d,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5 d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.
[목적]탐색건립인자궁내막이위증형광체외여재체모형적구체방법.[방법]채용증강형록색형광단백선병독(Ad-eGFP)분별전염원대배양인자궁내막선상피세포화기질세포(세포전염주사법,즉방법1)화자궁내막조직괴(조직전염주사법,즉방법2),비교량충체외전염적차이;연후채용방법1장록색형광선병독전염후적선상피세포화기질세포주입라서피하형성형광병조,병차여방법2상비교,관찰직지건모후25 d,계산병조형광성모솔여병조형광존재시간,병급여조직감정.[결과]방법1여2건모후5 d량충방법병조형광양성솔분별체88.9%화22.2%,P=0.015;병조체외형광존재시간분별위(12±8)d여(7±4)d.병리조직학HE염색화면역형광공동감정현시병조래원우인자궁내막.[결론]응용Ad-eGFP전염후인자궁내막선상피세포화기질세포혼현액주사가성공건립라서피하인자궁내막이위증활체형광관찰모형.동시방법성모솔고,체외형광존재시간교장.
[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 immunofluoresceney.[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