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目的 :通过对几种不同抗冻剂组成、不同降、复温条件的深低温保存方法对角膜内皮影响 ,探讨简便、有效的保存方法。方法 :将猪角膜分为A～E五组 ,分别给予或不给硫酸软骨素 (CS) ,E组不加入聚乙烯吡咯烷酮 (PVP) ,降温速率设置为 2步或 5步 ,37℃或 6 0℃水浴复温 ,保存 15天～ 3月 ,分别取出进行角膜内皮细胞光镜及电镜下观察。结果 :冷冻保护剂中添加 2 5 %CS、 5步降温可以有效提高保存效果 ,冷平衡由 4步改为 2步其保存效果基本不变 ,所保存的角膜内皮细胞形态接近正常 ,内皮细胞密度、存活率不随保存时间延长而发生明显下降。结论 :在冷冻保护剂中添加 2 5 %CS ,采用 5步降温的“二步法”方法简便 ,保存效果良好 ,在实践应用中有推广价值
OBJECTIVE: To investigate the effect of cryopreservation on the corneal endothelium by several kinds of cryoprotectants with different compositions of cryoprotectants, different hypothermia and rewarming conditions, and to discuss the simple and effective preservation methods. Methods: The porcine corneas were divided into five groups A to E, respectively, with or without chondroitin sulfate (CS), group E without polyvinylpyrrolidone (PVP). The cooling rate was set at 2 or 5 steps, 37 ℃ or 6 0 ℃ water bath rewarming, save 15 days to March, were removed corneal endothelial cells were observed by light microscopy and electron microscopy. Results: Adding 5% CS into the cryoprotectant could effectively improve the preservation effect by 5 steps of cooling. The cold balance was changed from 4 steps to 2 steps, the preservation effect was basically unchanged, the shape of corneal endothelial cells preserved was close to normal, the density of endothelial cells , Survival rate does not decline significantly with the preservation of time. CONCLUSION: The two-step method with 25% CS in cryoprotectant and 5-step cooling is simple and convenient, which has good preservation effect and has promotion value in practical application