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目的 探讨在羊膜上培养角膜缘上皮细胞 (limbal epithelial cells,LEC)的合适方法。方法 切取大小约 2 mm× 2 mm、厚约 2 0 0 μm含有完整上皮细胞的兔角膜缘组织 ,剪切成 4个 1 mm× 1 mm小的组织块 ,以羊膜为底物分别使用组织块培养法、组织块培养后传代培养法和组织块经消化酶处理后培养法培养兔 L EC,通过倒置显微镜、细胞组织学和扫描电镜观察细胞生长情况。结果 使用上述 3种方法培养的 L EC均可在羊膜上形成密集单层。细胞组织学检查显示在羊膜上培养的 LEC尚可形成多层。扫描电镜观察 LEC立体感强、细胞表面微绒毛丰富。但以组织块经消化酶处理后培养法培养 L EC较为快速。结论在羊膜上上述 3种方法都可用于 LEC的培养 ,但以组织块经消化酶处理后培养法更为实用
Objective To explore a suitable method for culturing limbal epithelial cells (LECs) on amnion. Methods The corneal limbal tissue of rabbit with intact epithelial cells about 2 mm × 2 mm in size and about 200 μm in thickness was excised and cut into 4 small pieces of 1 mm × 1 mm. The amniotic membrane Culture method and tissue culture were used to culture rabbit LEC after subculture and tissue culture with digestive enzyme. Cell growth was observed by inverted microscope, cell histology and scanning electron microscopy. Results LECs cultured using the above three methods all formed dense monolayers on the amniotic membrane. Histological examination showed that LEC cultured on amnion could form multiple layers. Scanning electron microscopy LEC stereoscopic strong cell surface microvilli is rich. However, culturing LEC with tissue culture after digestive enzyme treatment is faster. Conclusion The above three methods can be used for the culture of LEC in amniotic membrane, but it is more practical to use culture method after digestion enzyme treatment