摘要
目的 探讨人角膜缘干细胞 (stem cell,SC)体外培养方法并观察其生物学特性。方法 应用消化培养法对人角膜缘组织进行原代培养 ,记录其生长特性。待细胞生长至80 %汇合时 ,分别采用针对 6 4KD角蛋白的 AE5、针对一组酸性角蛋白的 AE1单克隆抗体 ,抗增殖细胞核抗原单克隆抗体 (anti- proliferating cell nuclear antigen,PCNA) ,针对5 0 KD烯醇酶特异的单克隆抗体 4G 10 .3,通过间接免疫荧光细胞化学染色对培养细胞进行鉴别。结果 人角膜缘组织应用消化培养法在 6～ 7d达到 80 %汇合状态 ,应用间接免疫细胞化学染色 ,AE1、AE5染色 ,胞浆染色阳性。 PCNA染色、细胞核染色阳性 ,4G 10 .3亦可见阳性结果。结论 应用消化培养法可成功地对人角膜缘组织进行原代培养 ,培养后细胞既可保持角膜上皮特性 ,又含有具有强增殖潜力的原始细胞 。
Objective To select culture technique of human limbal stem cell in vitro and observe its biological characteristics.Methods Human limbus was cultured primarily by enzymatic digestion culture method.When 80% confluence was reached,the cultured cell was differentiated by indirect immunochemically staining by a group of monoclonal antibodies including AE1,which recognized a group of acidic keratins,AE5,which recognized 64KD keratin,anti proliferating cell nuclear antigen,and 4G 10.3,which recognized 50KD enolase.Results Human limbus can be cultured successfully in this way .Eighty percent confluence was reached on 6th～7th day.All of the indirect immunochemically staining results were positive.Conclusion The cultured epithelial cells which are taken from limbus including stem cells can maintain their characteristics of cornea.There are some progenital cells with high proliferative potency.The cultured cells can be transplanted in clinic to treat severe ocular surface disease.
出处
《眼科新进展》
CAS
2001年第4期231-234,共4页
Recent Advances in Ophthalmology
基金
山东省医药卫生九五攻关课题资助 (970 917)
