SD大鼠骨组织冷冻预处理温度的选择

Selecting of Optimal Temperature for Frozen of SD Rats Bone

目的 探讨冷冻预处理温度对骨组织超微结构的影响 ,为吻合血管同种异体骨保存移植提供依据。 方法 以 1 5 %二甲基亚砜 (DMSO)为低温保护剂 ,采用两步冷冻步骤 ,对 SD大鼠骨组织进行 - 4 5 ,- 5 0 ,- 5 5 ,- 6 0 ,- 6 5和 - 70℃冷冻预处理 ,- 1 96℃液氮保存 1周 ,复温后光镜和透射电镜观察。 结果 温度 - 5 5℃冷冻预处理和液氮保存的大鼠骨组织超微结构保持良好 ,其余温度组的骨细胞轻度水肿 ,线粒体肿胀 ,嵴破坏 ,细胞核有固缩现象 ,部分骨细胞变性明显 ,甚者细胞膜不连续。 结论 骨组织冷冻预处理温度在 - 5 0～ - 6 0℃之间 ,- 5 5℃是最佳温度值 。

Objective To investigate the effect of preliminary treatment at a range of temperature in the ultrastructure of frozen bone of SD rat. Methods Two steps were adopted, firstly the samples were treated with frozen protection reagent(15% DMSO) and there were frozen at -45, -50, -55, {-60}, -65 and -70 ℃ for preliminary temperature treatment, then samples were stored at -196 ℃ liquid nitrogen for one week. The cell morphological changes were studied by using light mioroscopy and transmissor electron microscopy after rewarming. Results Pretreatment temperature of -55 ℃ were the optimal temperature for preservation the ultrastracture of bone cell was nearly normal, other temperature groups show: bone cell was slight edema, the mitochondria obvious dropsy and the mitochondrial crista of cytoplasm were destruction, some bone cell appeared karyon pyknosis and cell membrane disrupted. Conslusion The preliminary treatment temperature of frozen of bone tissue is -55^-60 ℃, {-55 ℃} may be the optimal preconditioning frogen temperature.;