以干燥脱水法保存的异种角膜基质为载体构建人工生物角膜上皮组织

目的:观察以干燥脱水法保存的鸵鸟角膜基质为载体构建人工生物角膜上皮组织的生物学特性。方法:采用组织块培养法获得新西兰大白兔角膜缘干细胞,经胰蛋白酶消化法获得细胞,种植于干燥脱水法保存的鸵鸟角膜板层基质上,采用气液界面培养法进行培养,通过倒置显微镜、透射电子显微镜、荧光显微镜观察其形态学、生长特点,超微结构及免疫学特征。结果:在干燥脱水法保存的鸵鸟角膜基质上种植兔角膜缘干细胞,接种72h后,细胞形成单层,移置气液交界面后继续培养7～10d,逐渐形成复层。经光镜、透射电镜、及免疫学检测显示其具有角膜上皮组织的生物学特性。结论:兔角膜缘干细胞能够在干燥脱水法保存的鸵鸟角膜基质载体上生长,并可形成复层,基本具有正常角膜上皮细胞的形态、超微结构和生物学特性。

Constructing artificial biologic cornea epithelial tissue on the carrier of heterogenic corneal stroma stored with desiccation and dehydration method

AIM:To observe the biologic features of the artificial biologic cornea epithelial tissue constructed on the carrier of heterogenic corneal stroma stored with desiccation and dehydration method.·METHODS:Limbal stem cells of New Zealand white rabbits were obtained with tissue block culture method.The cells gotten from trypsin digestion method were implanted on heterogenic corneal stroma stored with desiccation and dehydration,and cultured by air-liquid culture method.The morphology,growth,ultrastructure and immunology characteristics were observed under inverted microscope,transmission electron microscopy and fluorescence microscope.·RESULTS:After 72 hours,limbal stem cells implanted on heterogenic corneal stroma formed single layer.Seven to ten days after transplanted to air-liquid interface,the cells formed multi layer,which had biological characteristics of corneal epithelium tissue under inverted microscope,transmission electron microscopy and fluorescence microscope.·CONCLUSION:Limbal stem cells of rabbits can grow on the carrier of heterogenic corneal stroma stored with desiccation and dehydration method,and can form multi layer.The cells have the shape,ultrastructure and biological characteristics of corneal epithelium tissue.·