揭膜法培养兔角膜内皮细胞的改进

目的探讨培养兔角膜内皮细胞的最佳方法。方法在手术显微镜下撕下兔角膜后弹力层及内皮细胞层并使之完整覆盖于盖玻片上,避免其卷曲和折叠并剪切成均匀小块,使内皮面向上置于培养瓶中进行培养。结果未加入任何促细胞分裂剂的情况下,细胞在体外生长良好,角膜内皮细胞很快自组织片迁移生长,1周后组织块周围长出细胞晕,2周内铺满瓶底,可以快速获取角膜内皮细胞。结论本法培养的内皮细胞不需用酶,具有组织片无折叠,贴壁更牢固,细胞更易移行、增殖等优点,可为角膜内皮的研究提供较多的纯内皮细胞。

Improved stripping film emulsion method for cultivation of corneal endothelial cell of rabbits

Objective To explore an improved method to culture corneal endothelial cells.Methods Through surgical microscope,Descemet membrane and endothelial cell layer were isolated and overlaid on cover glasses,then dissected and seeded in culture containers with endothelial cell layer surface upwards.Results The cells grew well without adding any mitogen in the tissue culture medium.The corneal endothelial cells grew and were confluent within two weeks,the corneal endothelial cells could successfully obtain.Conclusion Enzyme is not required in this method;moreover,the method has many advantages,such as unfold tissue pieces,firm attachment,easy migration,and fast proliferation.With this technique,sufficient quantities of normal live cells become available for research of corneal endothelium.