新生大鼠视网膜干细胞分离培养及其生物学特性  

The biological characteristics of isolation and culture of retinal stem cell in neonatal rat

作　　者：王能[1] 栾文斌 李茂[1] 赵海洋[1] 项平[1] 

机构地区：[1]蚌埠医学院第一附属医院中心实验室,组织移植安徽省重点实验室,安徽蚌埠233004 [2]山东省蓬莱市中医院眼耳鼻喉科,265600

出　　处：《蚌埠医学院学报》2014年第3期281-283,共3页Journal of Bengbu Medical College

基　　金：安徽省教育厅自然科学研究重点资助项目(KJ2010A241)

摘　　要：目的:探讨新生大鼠视网膜干细胞的分离培养及其生物学特性。方法:采用机械酶-分离法体外扩增培养SD大鼠视网膜干细胞,观测细胞增殖能力,绘制CCK-8检测生长曲线;诱导细胞分化,免疫组织化学检测细胞巢蛋白、溴脱氧尿核苷、微管缔合蛋白-2、神经胶质原纤维酸性蛋白、胸腺糖蛋白1.1的表达。结果:原代视网膜干细胞形成大量的细胞球悬浮生长,传代后的细胞也可以形成细胞球,呈线性生长,巢蛋白和溴脱氧尿核苷表达阳性;诱导分化后的细胞部分原纤维酸性蛋白、微管缔合蛋白-2、胸腺糖蛋白1.1表达阳性。结论:新生SD大鼠视网膜干细胞可在体外分离培养和扩增,表达干细胞特性,并可向视网膜节细胞分化。Objective：To observe the results of isolation and culture of retinal stem cell in the neonatal rat and investigate their biological characteristics. Methods ： The mechanical enzyme separation in vitro amplification of rat retinal stem ceils was done, the capacity of cell proliferation and growth curve of CCK-8 detection were observed;cell differentiation was induced,then the expressions of nestin,bromodeoxyuridine, microtubule-associated protein-2, glial fibrillary acidic protein and thymus glycoprotein-1. 1 in cells were detected by immunocytochemical method. Results：The primary retinal stem ceils formed a large number of cells in the ball suspended growth, the passaged ceils could also form a ball of cells, and behaved as linear growth, the nestin and bromodeoxyuridine expressions were positive; glial fibriltary acidic protein, microtubule-associated protein-2, thymus glycoprotein-1.1 were positively expressed in part of cells after induction of differentiation. Conclusions： The neonatal SD rat retinal stem cells can be isolated, cultured, and amplified in vitro, expressed the characteristics of stem ceils, it can differentiate into retinal ganglion cells.

关 键 词：干细胞 视网膜 细胞培养技术 细胞分化 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]