摘要
目的：本研究利用组织工程原理和干细胞技术，构建尿道组织，修复兔尿道全段缺损。对细胞与细胞外基质材料在体外的复合方法，复合细胞材料修复尿道缺损的过程及其与单纯细胞外基质材料修复效果的差异，种子细胞在植入机体后的命运转归进行初步探讨，为进一步研究奠定基础。同时初步探讨利用BMSC-FG作为填充材料(Bulking Agent，BA)治疗压力性尿失禁及膀胱输尿管返流的可行性。
方法：①选择兔膀胱尿路上皮细胞、兔膀胱平滑肌细胞及兔骨髓间充质干细胞(BMSC)作为种子细胞，采用相应的培养及纯化策略，获取满足实验要求的种子细胞。并对两种可行的上皮细胞培养方案进行对比，选择更好的培养方法。利用细胞计数及MTT法对三种细胞在体外的生长增殖情况进行研究。利用脂质体转染对兔BMSC进行绿色荧光蛋白基因转染，试图获得稳定表达。②选择两种常用的泌尿外科组织工程材料——同种膀胱细胞外基质BECM和猪小肠粘膜下层SIS分别与兔BMSC及兔尿路上皮细胞进行复合培养，在体外构建尿道工程组织，对细胞在材料表面的分布及增殖情况进行研究。③分别利用复合有兔BMSC及尿路上皮细胞的兔膀胱细胞外基质对长度为1.5—2.0cm的雄性兔悬垂部全长尿道缺损进行原位修复实验，并与单纯BECM修复进行对比，32只动物分
Objective: To construct engineering urethral tissue in vitro using technique of Tissue Engineering and stem cells for repairing the urethral defect of a rabbit model. Looking for better ways for construction of urethra in vitro, and comparing the different between involve seeded matrices and unseeded matrices in the repairing process. To elucidate the destiny of the seeding cells in vivo and the possibility of using BMSC with fibrin glue as a bulking agent to treat stress incontinence and vesico-ureteral reflux.
Methods: (1) Bladder urothelium, bladder smooth muscle cells and bone marrow mesenchymal stem cells (BMSC) were choose as seed cells, primary culture and purify system were build to proliferate the seed cells. Two culture system (method 1 and method 2) were built for urothelium and one was choose to culture them in posterior studies. The proliferation characteristic of these cells were studied using cell counting and MTT Cell Proliferation Assay. To modify the BMSC, a liposome based plasmid transfection system was used to transfect BMSC, and its proliferation condition was studied after transfection. (2)Seeding BMSC and urothelium on the BECM and SIS was
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