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子宫内膜异体移植建立小鼠子宫内膜异位症模型的研究

发布时间:2018-02-10 03:07

  本文关键词: 子宫内膜异位症 GFP小鼠 sp细胞 出处:《福建医科大学》2014年硕士论文 论文类型:学位论文


【摘要】:背景:子宫内膜异位症的“干细胞学说”认为子宫内膜干细胞随月经逆流种植腹腔是内异症形成的关键,,但至今未有实验证据证明。 目的:探索小鼠子宫内膜是否存在干细胞;利用带有绿色荧光蛋白基因(GFP)的小鼠子宫内膜细胞或组织建立ICR小鼠子宫内膜异位症模型。 方法:1.分离和收集含GFP基因小鼠的子宫内膜细胞,经流式分选出SP细胞。 2.将收集到的GFP内膜细胞及组织移植于小鼠子宫浆膜及盆腔损伤处,实验分组:1)移植子宫内膜细胞组2)移植子宫内膜细胞与骨髓间充质细胞共培养组3)盆腔腹膜损伤后,直接移植消化的子宫内膜微组织组4)盆腔腹膜损伤3周后,移植消化的子宫内膜微组织组。继续饲养4-6周,荧光显微镜下直接观察病灶情况,并进行HE染色、免疫组化。 结果:1.新鲜消化的单细胞用Hoechst染色,经流式细胞仪分选出SP细胞群占1.25-2.29%。 2.含GFP基因的新鲜子宫内膜微小组织碎片移植于普通ICR小鼠可获得异位生长,经预损伤处理的小鼠较新鲜损伤移植的小鼠模型有更高的移植物生长率,分别为100%(12/12) VS37.5%(3/8),移植生长物GFP标记物阳性。 结论:小鼠子宫内膜细胞中含有SP细胞;微组织移植比细胞移植易形成移植灶,预损伤可以提高子宫内膜异体移植的成功率。
[Abstract]:Background: the stem cell theory of endometriosis suggests that endometrial stem cell implantation with menstrual countercurrent is the key to the formation of endometriosis, but there is no experimental evidence to prove it. Aim: to explore the existence of stem cells in mouse endometrium and to establish a model of endometriosis in ICR mice by using the cells or tissues of mouse endometrium with green fluorescent protein gene (GFP). Methods the endometrial cells of mice with GFP gene were isolated and collected. Sp cells were separated by flow cytometry. 2. The collected GFP endometrial cells and tissues were transplanted to the uterine serous and pelvic injury sites of mice. The experimental groups were divided into 2 groups: 1) transplanted endometrial cells group (2) transplanted endometrial cells and bone marrow mesenchymal cells co-cultured group 3) after pelvic peritoneal injury. After 3 weeks of pelvic peritoneal injury, the grafted endometrium microtissue group was kept for 4-6 weeks. The lesions were observed under fluorescence microscope and stained with HE and immunohistochemistry. Results the freshly digested single cells were stained with Hoechst, and SP cells were isolated by flow cytometry. The percentage of SP cells was 1.25-2.29. 2.Transplantation of fresh endometrial microtissue fragments containing GFP gene into normal ICR mice could obtain ectopic growth, and the predamaged mice had higher graft growth rate than the fresh injured mice. VS 37.5% were positive for GFP markers of transplanted growth. Conclusion: there are SP cells in mouse endometrial cells, microtissue transplantation is easier than cell transplantation to form transplantation focus, preinjury can improve the success rate of endometriosis transplantation.
【学位授予单位】:福建医科大学
【学位级别】:硕士
【学位授予年份】:2014
【分类号】:R711.71;R-332

【参考文献】

相关期刊论文 前1条

1 胡菲菲;廖联明;刘嘉茵;;产后小鼠子宫内膜边缘群干/祖细胞的分离纯化[J];生殖医学杂志;2008年04期



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