兔舌黏膜细胞构建组织工程化角膜上皮的实验研究  

Experimental study on amniotic membrane as the substrate for cultivating rabbit lingual epithelial cells in vitro

作　　者：顾正宇[1] 廖荣丰[1] 

机构地区：[1]安徽医科大学第一附属医院眼科,合肥230022

出　　处：《临床眼科杂志》2012年第6期546-548,共3页Journal of Clinical Ophthalmology

摘　　要：目的探讨兔舌黏膜上皮细胞以去上皮羊膜为载体,体外构建组织工程角膜上皮的可行性和方法,试图为眼表重建寻求另一种来源丰富的有效组织工程化角膜移植材料。方法胰酶消化去除羊膜上皮细胞,保留基底膜作为载体,将兔舌黏膜上皮细胞种植于去上皮羊膜基底膜进行体外培养扩增。对培养获得的复合上皮片进行形态观察、常规病理切片HE染色和透射电镜观察。采用AE1/AE3单克隆抗体鉴定复合上皮片中有无角蛋白表达。结果兔舌黏膜上皮细胞在去上皮羊膜基底膜面能黏附生长并增殖,体外培养2～3周左右即可在羊膜上汇合成片并分层。形成的复合上皮片由羊膜基底膜和2～3层较扁平的上皮细胞组成,细胞之间可见桥粒连接,细胞表达角蛋白。结论兔舌黏膜上皮细胞以去上皮羊膜为载体,经体外培养扩增获得的复层组织,初具角膜上皮的雏形结构。Objective To investigate the cultivation method and feasibility of rabbit lingual epithelial cells planted on anmiotic membrane （AM） as a substrate and try to find out satisfactory methods and rich material for ocular surface re- construction. Methods Rabbit lingual epithelial cells （ EC ） were planted on denuded AM. The resulted sheet was exam- ined by inverted microscope, HE staining pathological section and electron microscopy. The sheet was also immunohisto- chemically labeled with AE1/AE3 monoconal antibody of Pan cytokeratius. Results Rabbit lingual ECs could stick and confluence on denuded AM. After 2 - 3 weeks in culture, two to three layers of stratification lingual epithelial cells had been established with the presence of Pan CK. EM revealed that numerous desmosomal junctions were among the multiple layers of lingual ECs. Conduslon Tissue engineering corneal epithelium similar to normal rabbit corneal epithelium can be obtained by implanting rabbit lingual ECs on denuded AM.

关 键 词：组织工程 羊膜 舌黏膜上皮 

分 类 号：R318.08[医药卫生—生物医学工程]