三种不同冷冻保护剂对小鼠附睾冷冻效果的比较

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)%(P0.05);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。

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

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 .