羊膜对山羊角膜缘上皮细胞培养影响的研究

摘要

比较完整羊膜和去上皮羊膜、不同冻存时间的羊膜对山羊角膜缘上皮细胞体外培养的影响,筛选出羊膜最适处理方法.将山羊角膜缘组织块分别接种于完整羊膜、去上皮羊膜、新鲜去上皮羊膜、冻存30 d去上皮羊膜、冻存90 d去上皮羊膜、冻存180 d去上皮羊膜上,比较在完整羊膜和去上皮羊膜及不同冻存时间羊膜上角膜缘上皮细胞的生长特性和组织结构.试验结果显示,在去上皮羊膜上角膜缘组织块贴壁和细胞增殖能力较完整羊膜强,角膜缘上皮细胞在2组羊膜上均能形成多层结构,但完整羊膜上形成的组织表层角质化较严重；在冻存30 d羊膜上角膜缘组织块贴壁和细胞增殖能力较新鲜羊膜、冻存90 d羊膜、冻存180 d羊膜强,角膜缘上皮细胞在各组羊膜上均形成多层结构,其结构无明显差异.结果表明,冻存30 d去上皮羊膜是构建组织工程人工角膜最适的羊膜载体材料.%The differrnces of goat corneal limbal epithelial cells cultured respectively on intact amniotic membrane, denuded amniotic membrane and cryopreserved denuded amniotic membrane treated in different freezing time were assessed to find a optimum treatment method of amniotic membrane in this study. Limbal explant were cultured on the intact amniotic membrane, denuded amniotic membrane, uncryopreserved denuded amniotic membrane, 30-day cryopreserved denuded amniotic membrane, 90-day cryopreserved denuded amniotic membrane and 180-day cryopreserved denuded amniotic membrane respectively. The contrasts were performed among the growth characteristics and morphologies of corneal limbal epithelial cells seeded on different treated amniotic membrane. The results showed that the adherence and proliferation of cells cultured on denuded amniotic membrane were better than that of cells cultured on intact amniotic membrane. The cells cultured on both of the amniotic membranes could grow and form multilayer cellular structure. However, the keratinization of superficial cells cultured on the intact amniotic membrane was more than that of superficial cells cultured on denuded amniotic membrane. The adherence and proliferation of cells cultured on 30-day cryopreserved denuded amniotic membrane were better than that of cells cultured on uncryopreserved denuded amniotic membrane, 90-day cryopreserved denuded amniotic membrane and 180-day cryopreserved denuded amniotic membrane. The cells on all different treated amniotic membrane could grow and form multilayer cellular structure. However, no significant differences were found among the cellular structure of the different treated amniotic membrane. The 30-day cryopreserved denuded amniotic membrane was the optimum choice which could be used for reconstructing tissue engineering artificial cornea.