【摘要】 目的:观察5-乙炔基-2’脱氧尿嘧啶核苷(Edu)标记的人脐带间充质干细胞(huc MSCs)在小鼠慢性输卵管炎模型中的分布与定位。方法:用Mo Pn衣原体感染液阴道内注射建立小鼠输卵管炎模型,体外培养人脐带间充质干细胞,用10μmol/L Edu培养液对huc MSCs孵育不同时间(0小时、24小时、48小时、72小时)进行标记,同时检测标记组和未标记组细胞增殖和凋亡情况。感染后4周随机分成huc MSC治疗组及PBS对照组,感染后8周,亚甲蓝通水试验观察小鼠输卵管有无积水、阻塞等慢性输卵管炎表现,病理切片HE染色观察组织形态学改变,荧光共聚焦显微镜下观察Edu标记的huc MSCs移植治疗在小鼠输卵管中的分布与定位情况。结果:实验成功建立了小鼠慢性输卵管炎模型,Edu标记前后的huc MSCs的形态无明显差别,均为成纤维样细胞,细胞增殖及凋亡未见明显影响,细胞生长状态良好。10μmol/L Edu标记huc MSCs孵育48小时即可获得良好的阳性细胞标记率(45.53±1.19)%。Edu标记阳性的huc MSCs主要分别在输卵管间质中。huc MSCs治疗能够缓解输卵管积水、阻塞情况,减轻慢性炎症。结论:在小鼠慢性输卵管炎模型中可观察到Edu标记的阳性huc MSCs,Edu标记技术可用于huc MSCs在小鼠慢性输卵管炎模型中定位的研究。更多还原

【Abstract】 Objective: To observe the distribution and localization of 5-ethynyl-2’deoxyuridine( Edu) labeled human umbilical cord mesenchymal stem cells( huc MSCs) in mouse model of chronic salpingitis.Methods: Chronic salpingitis mouse model was established by intravaginal injection of Mo Pn chlamydia. The huc MSCs were cultured in vitro and incubated with 10 μmol/L Edu medium for different time( 0 h,24 h,48 h,72 h),and the proliferation and apoptosis of the labeled and unlabeled cells were simultaneously detected. The cells were randomly divided into huc MSC treatment group and PBS control group at 4 weeks after infection.And after 8 weeks,methylene blue water test was conducted to observe whether the mouse occurred hydrosalpinx,oviduct obstruction or other chronic salpingitis manifestations.Pathological sections of HE staining was used to observe the histomorphological changes,and fluorescence confocal microscopy was used to observe the distribution and localization of Edu labeled huc MSCs in mouse oviduct.Results: The model of chronic salpingitis in mouse was established successfully.There was no significant difference in the morphology of huc MSCs between before and after Edu labeling. All of them were fibroblast-like cells,and no significant effect was observed in the cell proliferation and apoptosis.The huc MSCs labeled 10 μmol/L Edu for 48 hours can obtain good positive cell labeling rate( 45. 53 ± 1. 19) %. The Edu-positive huc MSCs were mainly distributed in tubal interstitial,and huc MSCs transplantation could alleviate hydrosalpinx and oviduct occlusion,and reduce the chronic inflammation.Conclusions: Edu labeled huc MSC can be observed in the chronic salpingitis mouse.The Edu labeling technique can be used to study the location of huc MSCs in mouse with chronic salpingitis.更多还原