Objective To establish a convenient, economic and pragmatic method for rabbit corheal epithelial cells primary culture, and observe the biologic characteristics of growth in vitro. Methods The rabbit corneal epithelial cells were cultured by the entire corneal tissue - culture. The morphological characteristics of cultured cells were observed by inverted phase contrast microscope at various time periods respectively and the cultured cells were identified by immunohistochemical method. Results The rabbit entire cornea adhered in 2 days, the cells could be seen climbing out from the corneal limbus. The cells and nuclei were spherical and nuclei were located next to the central area or in the center; The epithelial cells were flat and polygonal and connected to each other into membrane in 3 days; Suspension of cell growth can be seen locally in 7 days of primary culture; The corneal epithelial cells grew vigorously and some cells were adhered and were flaky and polygonal on day 2 of passage 2. The cultured cells stained positively for monoclonal antibody pan cytokeratin by immunohistochemical method. Conclusion The entire corneal tissue - culture is a simple convenient and practicable method for culturing corneal epithelial cells which could be serially subcultivated.%目的 建立体外原代培养兔角膜上皮细胞简单经济实用的方法,并观察其体外生长的生物学特性.方法 采用全角膜组织培养法培养兔角膜上皮细胞并进行传代培养,通过形态学观察并应用免疫组织化学方法对细胞进行鉴定.结果 兔角膜上皮原代培养第2天,组织边缘有细胞游出,细胞和细胞核均呈圆形,胞核位于细胞中央或旁中央区；第3天,细胞呈扁平多边形,相互连接成片；兔角膜上皮原代培养第7天,局部可见悬浮细胞生长；兔角膜上皮细胞培养第2代第2天,细胞生长旺盛,部分细胞贴壁呈片状多边形.兔角膜细胞免疫组织化学显示广谱角蛋白单克隆抗体阳性.结论 全角膜培养角膜上皮细胞简单实用,可获得状态良好的连续传代扩增的细胞.
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