兔角膜内皮细胞的原代培养

Primary culture of endothelial cells of rabbit corneas

目的:探讨兔角膜内皮细胞的培养方法。方法:显微分离兔角膜后弹力层-内皮细胞层,采用贴壁法、贴壁消化法及两种消化法培养,观察每种方法细胞生长的特点。结果:4种方法培养的兔角膜内皮细胞均呈单层密集生长,形态为六角形或多边形,具有活体细胞的形态特征。培养周期最短约为1周。结论:将揭取的附有内皮细胞的后弹力层预培养1天后再进行消化培养的方法,可为实验研究提供有效的角膜内皮细胞培养方法。

Objective： To provide a sound basis for the experimental research by cultivation of endothelial cells from rabbit corneas. Methods：The Deseemet＇ s membrane with its endothelial covering was separated from the underlying stroma by microdisseetion. Different methods were used to isolate and cultivate corneal endothelial cells of rabbits in vitro and their characteristics were examined. Results： Cultured corneal endothelial cells of rabbits were hexagonal or polygonal and formed a compact monolayer. The cultured cells appeared to maintain the morphologic characteristics of endothelial cells in vivo. The Descemet＇ s membrane-endothelium from rabbits was cultured for one day at first ,and then incubated with trypsin and EDTA in D-Hanks solution. The shortest culture period was about a week with this method. Conclusions：The Descemet＇ s membrane with its endothelial covering can supply for cultivating the epithelial cells of corneas.