深低温保存角膜缘组织的结构和增殖活性研究

目的 探讨深低温保存材料角膜缘组织的结构和增殖活性。方法 应用常规切片，HE染色及电镜检查观察深低温保存4个月的角膜缘组织的结构变化。应用免疫组织化学染色法对6片深低温保存4个月的角膜缘材料在刚复温时及于培养液中放置6d后的增殖细胞核抗原(PCNA)表达进行检测，2片新鲜角膜缘材料作为对照。结果 深低温保存4个月的角膜缘材料基本保持了其形态结构的完整性。新鲜角膜缘材料PCNA呈弱阳性表达，6片深低温保存材料在复温时其PCNA呈阴性表达，但当置于培养液中6d后，其PCNA呈阳性表达。结论 深低温保存4个月的角膜缘材料结构完整，且保持其增殖活性，可用于角膜缘移植。

Structure and proliferation activity of limbus tissue after cryopreservation

ObjectiveTo explore the stucture and proliferative activity of limbus tissue after cryopreservation. MethodsThe structure of limbus tissue after cryopreservation for 4 months was observed using electron microscope and HE staining. Expression of PCNA in 2 fresh limbus epithelium,6 limbus epithelium cryopreserved for 4 months and limbus epithelium cultured in DMEM/F12 for 6 days after cryopreservation was detected with Mouse anti-human monoclonal antibody.ResultsThe structure of limbus tissues after cryopreservation maintained integrity. The PCNA was expressed in some cells of fresh limbus epithelium.No positive staining of PCNA was found in limbus epithelium of all 6 specimens cryopreserved for 4 months. However, when the limbus tissues cryopreservated were placed in the culture medium for 6 days,the expression of PCNA was found.ConclusionCryopreservation has no effect on structure of limbus tissue. There is active proliferation of cryopreservated limbus tissue in suitable environment and this method can be used to limbus transplantation.