摘要
探索简易安全的冻贮角膜技术。方法 :对经典的 Capella四步冻贮法的冷冻保护液、降温冷冻速率和复温方法 3项关键技术做了全面简化和改进。以台盼蓝和茜素红联合染色评价家兔和人尸角膜内皮细胞存活率 (ESR)及光镜下内皮细胞形态。结果 :优选出“简化四步法”冻贮技术 :以 2 0 %人血清白蛋白与 2 %、 4%、 6 %和 7.5 %二甲基亚砜配制 4种冷冻保护液 ;角膜分别在 4种液体中冷平衡 15 m in;在简易降温仪内先以 1～ 3℃ / min降温至 - 30℃ ,再以 3～ 10℃ / min降温至 - 80℃ ;液氮中贮存 ;40℃水浴中复温。本组 ESR (92 .6 % ,n=30 )与 Capella组 (93.4% ,n=30 )比较 ,差异无显著性 (P>0 .0 5 ) ,两组间内皮细胞形态相仿。结论
Objective:To search for a simple and safe technique for cryopreservation of corneas.Methods:The tests about the three key techniques:cryoprotective medium,freezing and thawing methods of Capellas method were comparatively studied.Dual staining with trypan blue and alizarin red was used to evaluate the endothelial morphology and survival rate(ESR) of the rabbits and human corneas.Results:Select“simplified four step”technique is as follows.Freshly excised corneas were incubated at 4°C for 15 minuts in each of four cryoprotective media which consisted of 2%,4%,6% and 7 5%(V/V) dimethylsulfoxide(DMSO) respectively and 20% human sera albumin.Corneas were frozen in QL simple cryofreezer at the rate of 1°～3°C/min to -30℃,then 3～10℃/min to -80℃.Corneas frozen in liquid nitrogen were thawed in 40℃ water bath in modified manipulation.There were no sigmificant differences in ESR between this group and Capellas group(p>0 05).Corneal endothelial morphology was similar under light microscopy Conclusion:“simplified four step”technique is simple and safe.
出处
《中国实用眼科杂志》
CSCD
2000年第6期350-352,共3页
Chinese Journal of Practical Ophthalmology
基金
九五全军卫生医药科研基金 (青年项目 )资助 !(Q-960 0 1 )
