组织块培养角膜缘上皮细胞的生物学特征观察  

Observation of biological characteristics of limbal epithelial cells by tissue inoculation

作　　者：刘兴华[1] 朱美玲[1] 王明丽[1] 陈逖[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：《安徽医科大学学报》2002年第4期262-264,共3页Acta Universitatis Medicinalis Anhui

基　　金：安徽省教育厅基金资助项目 (编号 :2 0 0 0jl10 3 )

摘　　要：目的　观察以人羊膜为底物组织块培养角膜缘上皮细胞的生物学特征。方法　取含有上皮细胞大小约 2mm×2mm、厚约 2 0 0 μm兔角膜缘组织 ,剪切成 4个小的组织块 ,用消化液消化至组织块变疏松后放在去除上皮细胞的羊膜上培养 ,每 2～ 3d用倒置显微镜观察 ,定期取羊膜进行细胞组织学、免疫组织化学及电镜检查。结果　角膜缘组织块在羊膜上培养 8d ,羊膜上培养的角膜缘上皮细胞出现汇合并部分成层 ,培养 16d ,细胞形成 3～ 4层 ,并很好地附着于下面的羊膜上 ,羊膜上培养的细胞经广谱角蛋白 (pan keratin ,PK)抗体和抗增殖细胞核抗原 (proliferatingcellnuclearanti gan ,PCNA)单克隆抗体免疫组织化学染色呈阳性反应。结论　羊膜作为底物组织块培养的角膜缘上皮细胞短期内能形成多层且具有较强增殖能力 。Objective To observe biological characteristics of limbal epithelial cells(LEC) by tissue inocula tion on amniotic membrane(AM). Methods Small (about 2 0 mm×2 0 mm× 200 μm) specimens of LEC were excised from the limbal regions in rabbits. Each of the specimens was cut into four tissues. The tissues were loosen by enzyme digestion and cultivated on acellular human AM. The cultivated LEC on AM were observed by inverted microscope every two or three days and examined by histology, immunohistochemical and electron microscope. Results A confluent primary c ulture of LEC was established on acellular human amniotic membrane on the 7th day. After 16 days of culture, the epithelial cells formed 3～4 layers and attached well to the underlying amniotic membrane. The cultured cells on AM were stained positive in Pan keratin antibody and anti PCNA. Conclusion The tissue cultivated LEC on AM can form multi layers and some of them can proliferate. They can also be transplanted.

关 键 词：角膜缘 细胞学 组织培养 羊膜 

分 类 号：R322.91[医药卫生—人体解剖和组织胚胎学]