小鼠阴道上皮细胞与猪脱细胞真皮基质-纤维蛋白凝胶构建组织工程阴道的初步研究  

Primary Experimental Study on Tissue Engineering Vagina Constructed by Mouse Vaginal Epithelial Cells and Porcine Acellular Dermal Matrix (PADM)-Fibrin Gel

作　　者：张明乐[1] 黄向华[1] 李雅钗[1] 张琳[1] 李亚楠[1] 朱志洁[1] 

机构地区：[1]河北医科大学第二医院,河北石家庄050000

出　　处：《实用妇产科杂志》2012年第8期639-642,共4页Journal of Practical Obstetrics and Gynecology

基　　金：河北省医学适用技术跟踪项目(编号:GL2011-36)

摘　　要：目的:初步探讨利用组织工程学技术构建组织工程阴道的方法及可行性。方法:将小鼠阴道上皮细胞以1×105个/ml的密度接种到纤维蛋白凝胶内并与猪脱细胞真皮基质(PADM)复合构成移植片,体外培养48小时后,将移植片埋植到小鼠脊柱两侧皮下。移植后2周、4周、8周和12周取材,对埋植后不同时间取出的移植片行HE染色和VanGieso(VG)染色,观察和评价移植片组织的生长状况;用上皮广谱角蛋白单克隆抗体AE1/AE3进行免疫组化染色,以证实移植片是否存在上皮组织。结果:光镜下观察,皮下埋植后2周,可见少量阴道上皮细胞,4周形成上皮,仅2～3层,8～12周上皮层数增加(约4～5层)。AE1/AE3免疫组化染色结果与HE染色结果一致。VG染色显示随埋植时间推移,PADM内逐渐有细胞长入,结构逐渐降解。结论:阴道上皮细胞与PADM-纤维蛋白构建的移植物体内移植后有血管长入,形成的上皮组织表型与正常阴道上皮表型相同。Objectives： To investigate the feasibility of tissue engineering vagina constructed by vaginal epithelial cells, fibrin gel and porcine acellular dermal matrix （PADM）. Methods：Cultured vaginal epithelial cells were suspended in fibrin in a concentration of 1×10^/ml and grafted onto PADM. Biologic attachment and growth of cultured mice vaginal epithelial cells were observed daily. After culturing 48 hours in vitro, the cell-seeded scaffolds were implanted subcutaneously lateral to the spine of the mice. The animals were sac- rificed 2, 4, 8 and 12 weeks after implantation. The tissue-engineered grafts were prepared for HE and Van Gieson （VG） staining to evaluate the status of various cell layer growth. Epithelial broad spectrum of mono- clonal antibodies of AE1/AE3 was used to detect the existence of epithelial cells. Results：Two weeks after transplantation, only a few vaginal epithelial cells could be observed under the light microscopy. Within 4 weeks 2 -3 layers of epitheliums could be observed. Within 8 - 12 weeks, epithelization was intensified and 4 -5 layers of epitheliums could be observed. AE1/AE3 staining had the same results with HE staining. VG staining indicated that the cellular ingrowth were accompanied by the degradation of the grafts. Conclusions： Cell seeded polymer scaffolds are able to form vascularized vaginal tissue in vivo, and that the phenotypic properties of the epitheliums are similar to those of normal vaginal tissues.

关 键 词：阴道重建 组织工程 细胞培养 猪脱细胞真皮基质 

分 类 号：R713.3[医药卫生—妇产科学]