论文部分内容阅读
目的 观察以人羊膜为底物组织块培养角膜缘上皮细胞的生物学特征。方法 取含有上皮细胞大小约 2mm×2mm、厚约 2 0 0 μm兔角膜缘组织 ,剪切成 4个小的组织块 ,用消化液消化至组织块变疏松后放在去除上皮细胞的羊膜上培养 ,每 2～ 3d用倒置显微镜观察 ,定期取羊膜进行细胞组织学、免疫组织化学及电镜检查。结果 角膜缘组织块在羊膜上培养 8d ,羊膜上培养的角膜缘上皮细胞出现汇合并部分成层 ,培养 16d ,细胞形成 3～ 4层 ,并很好地附着于下面的羊膜上 ,羊膜上培养的细胞经广谱角蛋白 (pan keratin ,PK)抗体和抗增殖细胞核抗原 (proliferatingcellnuclearanti gan ,PCNA)单克隆抗体免疫组织化学染色呈阳性反应。结论 羊膜作为底物组织块培养的角膜缘上皮细胞短期内能形成多层且具有较强增殖能力 ,可以用来移植
Objective To observe the biological characteristics of corneal limbal epithelial cells cultured with human amniotic membrane. Methods The rabbit corneal limbal tissue with a size of about 2 mm × 2 mm and a thickness of 200 μm was cut into 4 small pieces of tissue and digested with digestion solution until the tissue became loose and placed on the amniotic membrane with epithelial cells removed Culture, every 2 ~ 3d observed with an inverted microscope, amniotic membrane taken regularly for histology, immunohistochemistry and electron microscopy. Results The corneal limbal tissue was cultured on the amniotic membrane for 8 days. The limbal epithelial cells cultured on the amniotic membrane appeared confluent and partially stratified. After cultured for 16 days, the cells formed 3 ~ 4 layers and attached well to the amniotic membrane below. Of the cells were positive for immunohistochemical staining with monoclonal keratin (PK) antibody and monoclonal antibody against proliferating cell nuclear antigen (PCNA). Conclusion The limbal epithelial cells cultured in amniotic membrane as a substrate can form multiple layers in a short period and have strong proliferative capacity and can be used for transplantation