摘要
背景:有实验报道,碱性成纤维细胞生长因子能改变角膜内皮细胞形状、促进其分裂和趋化,刺激活体和体外角膜内皮细胞的损伤修复。目的:观察碱性成纤维细胞生长因子对猫角膜内皮细胞增殖的影响。设计、时间及地点:细胞形态学观察实验,2005-01/2006-12在青岛大学医学院附属医院中心实验室完成。材料:家猫购买自青岛大学医学院附属医院动物实验中心。方法:猫角膜内皮细胞原代培养,行NSE免疫组织化学染色鉴定细胞的类型、纯度及生理特性。传1代后,分别在培养液中加1,10,100μL的碱性成纤维细胞生长因子,继续卵化1~5d。主要观察指标:加药后第1,3,5天分别利用MTT法测定在490nm处的吸光度值来判断角膜内皮细胞增殖情况;同时应用倒置相差显微镜观察细胞形态及透射电镜检测细胞超微结构。结果:碱性成纤维细胞生长因子作用组加药后1,3,5d,MTT法测定猫角膜内皮细胞在490nm处的吸光度值明显高于对照组,其中10μg/L作用组差别最显著。结论:碱性成纤维细胞生长因子作用能促进猫角膜内皮细胞的增殖,相对有效浓度为10μg/L。
BACKGROUND: Previous experiments reported that basic fibroblast growth factor (bFGF) can change the shape of corneal endothelial cells, accelerate cell division and chemotaxis, and repair corneal endothelia cells injury in vivo and in vitro. OBJECTIVE: To investigate the effect of bFGF on cat corneal endothelial cells proliferation. DESIGN, TIME AND SETTING: The cell morphological observation was performed at the Central Laboratory of the Affiliated Hospital of Medical College of Qingdao University between January 2005 and December 2006. MATERIALS: Domestic cats were purchased from Experimental Animal Center of the Affiliated Hospital of Medical College of Qingdao University. METHODS: The corneal endothelial cells of cats were primarily cultured, and neurone specific enolase (NSE) immunocytochemistry staining was performed. After cell passage, bFGF (1, 10, 100μg/L) was added, continuously hatched for 1 5 days. MAIN OUTCOME MEASURES: MTT method was used to determine the proliferation of corneal endothelial cells; meantime, inverted phase contrast microscope and transmission electron microscope were used to observe the ultrastructure of cells. RESULTS: At days 1, 3 and 5 after bFGF was added, the absorbance value in 490 nm was significantly higher than that of control group, especially in 10 μg/L group. CONCLUSION: The bFGF can promote proliferation of cat corneal endothelial cells with effective concentration of 10 μg/L.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第20期3911-3914,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research