【摘要】 目的玻璃化法和慢速冷冻法深低温保存兔颈总动脉,比较两种保存方法对血管活性及形态结构的影响。方法无菌条件下切取兔颈总动脉,采用玻璃化法和慢速冷冻法在-196℃液氮中深低温保存2周,37℃水浴复温。新鲜血管作为对照,分别观察血管的大体、组织学、超微结构;同时对冷冻复温的血管采用组织块法培养并鉴定细胞,通过细胞计数描述细胞生长曲线,观察培养后血管的活性、细胞的再生及增殖能力。结果玻璃化组和慢速冷冻组动脉出现部分内皮细胞脱落,平滑肌细胞和成纤维细胞轻度肿胀、变性,前者损伤程度轻于后者。体外培养两组动脉的细胞分别于第6、7天自组织块内长出,细胞形态结构、生物学特性与新鲜组相似。生长曲线显示玻璃化组细胞的生长增殖状态优于慢速冷冻组,两组均差于新鲜动脉。将培养后的细胞分别进行免疫组织化学染色,经鉴定为平滑肌细胞。结论玻璃化法能较好地保持血管的形态结构和组织活性,保存效果优于慢速冷冻法。更多还原

【Abstract】 Objective To assess the effects of vitrification and slow freezing method on the morphology and viability of rabbit carotids.Methods Rabbit carotids were harvested and cryopreserved by vitrification and slow freezing method for 2 weeks,and then thawed slowly in water bath at 37 ℃.The fresh carotids were taken as controls.The morphological changes of all carotids were observed with the naked eye,optical microscope and electron microscope.The fresh and frozen-thawed arteries were cultured in vitro.Viability,regeneration and proliferation of the cells were observed and described by cellular growth curve.Results The morphological studies demonstrated that some of endothelium cells(ECs) fell off,some of the smooth muscle cells(SMCs) and fibroblast cells swelled or denaturalized after freeze-thawing procedure.Vitrification,however,was less detrimental to vessel structure than slow freezing method.The cells of vitrified carotids and frozen carotids grew out from tissues on the 6th and 7th day.The biological characteristics were similar to those of the fresh carotids.The growth curve showed that cell viability and proliferation were preserved perfectly in vitrification group compared to slow freezing group,but inferior to the fresh carotids.The cells cultured from all carotids were identified through immunohistochemical staining and proved to be SMCs.Conclusion Vitrification does less damage to the morphology and viability of vessels,so rabbit carotids preserved by vitrification are better than those preserved by slow freezing method.更多还原