三种不同冷冻保护剂对小鼠附睾冷冻效果的比较  

Comparison of the effect of three cryoprotectants on vitrification-cryopreservation of mouse epididymis

作　　者：李淼[1] 郁丽丽 张艺宝[3] 强苏静 刘丽均 徐平[1,2] 

机构地区：[1]中国科学院上海生命科学研究院,上海201615 [2]上海斯莱克实验动物有限责任公司,上海201615 [3]南京农业大学动物医学系,南京210095

出　　处：《中国实验动物学报》2014年第2期62-65,I0007,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：上海市科研计划项目(111409022700);国家科技支撑计划课题(2013BAK11B02)

摘　　要：目的：探讨三种不同冷冻保护剂对C57BL/6J小鼠附睾冷冻的效果。方法性成熟并交配过的6～8周龄C57BL/6J雄鼠附睾，分别用3种不同冷冻保护剂二甲基亚砜（DMSO）、丙二醇（PROH）、R18S3进行玻璃化冷冻、复苏和精子采集，观察比较三个实验组的复苏精子形态、存活率以及生殖能力。结果三组冷冻复苏分离后的精子形态完整，具有镰刀头状结构；荧光染色后PROH组精子存活率为（88.17±3.43）％，明显高于DMSO组：（61.17±10.65）％和R18S3组（16.83±6.49）％（ P＜0.05）；经辅助体外受精后均具有受精能力。获得的胚胎移植后可得到正常子代小鼠（DMSO∶PROH∶R18S3＝13∶8∶17）。结论3种冷冻保护剂均适用于C57BL/6J小鼠附睾冷冻，但PROH冷冻效果较好。Objective To compare the effect of three cryoprotectants on vitrification-cryopreservation of C57BL/6J mouse epididymis.Method Epididymises from 6-8-week old male C57BL/6J mice （age） were cryopreserved using DMSO, PROH and R18S3 cryoprotectant solution and thawed , respectively.The morphology of sperm from thawed epididy-mis, the rate of post-thawing survival and reproductive capacity were determined to evaluate the freezing efficiency of the three cryoprotectants .Results The sperms from thawed epididymis of the three groups were all well-preserved structural-ly.The survival rate of sperms was 88.17%, 61.17% and 16.83% in the PROH, DMSO and R18S3 cryoprotectant solu-tions, respectively, and there were significant differences between the three cryopreservation groups （P〈0.05 for all）. The number of pups from the DMSO , PROH and R18S3 groups were 13, 8 and 17 mice after ICSI and embryo implantation , respectively.Conclusions All the three cryoprotectants DMSO , PROH and R18S3 solutions are suitable for vitrification-cryopreservation of C57BL/6J mouse epididymis.But PROH is the preference of these cryoprotectant solutions .

关 键 词：小鼠附睾 冷冻保护剂 冷冻效果 

分 类 号：Q95-33[生物学—动物学]