【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。更多还原

【Abstract】 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 R18S3cryoprotectant solution and thawed,respectively. The morphology of sperm from thawed epididymis,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 structurally. The survival rate of sperms was 88. 17%,61. 17% and 16. 83% in the PROH,DMSO and R18S3cryoprotectant solutions,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 R18S3groups were 13,8 and 17 mice after ICSI and embryo implantation, respectively. Conclusions All the three cryoprotectants DMSO,PROH and R18S3solutions are suitable for vitrificationcryopreservation of C57BL /6J mouse epididymis. But PROH is the preference of these cryoprotectant solutions.更多还原