Rendal E, Santos M V Martínez, Rodriguez M, Sánchez J, Segura R, Matheu G, Filgueira P, Pértega S, Andión C
Cryobiology Unit, Complejo Hospitalario Juan Canalejo, La Coruña, Spain.
Transplant Proc. 2004 Dec;36(10):3283-7. doi: 10.1016/j.transproceed.2004.10.057.
The objective of this study was to analyze the importance of the quality of the vascular segment to be cryopreserved and the influence of storage in a gas phase, a liquid phase, or after accidental immersion in liquid nitrogen. In addition, we investigated the effects of rapid versus slow thawing on the occurrence of fractures and changes in the structure of the vessel wall. The tissue sources were whole thoracic and abdominal aortas from 15 pigs. Each aorta was cut into equal segments and randomly assigned to each study group. One segment of fresh unfrozen aorta of the same size was used as a control. The samples were cryopreserved using a programmed apparatus. After 2 weeks the arterial segments were thawed rapidly or slowly. A great variation in the results was obtained depending on the quality of the control. Although endothelial cells were better preserved in the liquid phase, the internal elastic lamina and elastic lamelli showed better preservation and fewer microfractures in the gas phase. The internal elastic lamina showed a greater number of microfractures when an accidental immersion in liquid nitrogen had taken place. Furthermore, better preservation of the structure of the vascular segment was observed with a slow thawing method. In general, the conditions of storage and the method of thawing seem to damage the structure of vascular segments. It is necessary to use a severe protocol of donor and vascular segment selection to optimize the post-thaw quality of the cryopreserved samples.
本研究的目的是分析待冷冻保存的血管段质量的重要性以及气相、液相储存或意外浸入液氮后的影响。此外,我们研究了快速解冻与缓慢解冻对血管壁骨折发生率和结构变化的影响。组织来源为15头猪的全胸主动脉和腹主动脉。将每根主动脉切成相等的节段,并随机分配到每个研究组。取一段相同大小的新鲜未冷冻主动脉作为对照。使用程序控制的仪器对样本进行冷冻保存。2周后,将动脉节段快速或缓慢解冻。根据对照的质量,结果有很大差异。虽然液相中内皮细胞保存较好,但气相中内弹性膜和弹性层板保存较好且微骨折较少。当意外浸入液氮时,内弹性膜出现更多的微骨折。此外,缓慢解冻方法观察到血管段结构保存更好。一般来说,储存条件和解冻方法似乎会破坏血管段的结构。有必要采用严格的供体和血管段选择方案,以优化冷冻保存样本的解冻后质量。