小鼠前向运动精子分离采集技术改良

Improved Method for Isolating and Collecting Mouse Forward Motile Sperm

目的提高从小鼠附睾采集的精子中前向运动精子的比例。方法设计"双舱培养皿"和"双舱浮游法(B法)"采集小鼠精子,并与当前普遍采用的"常规浮游法(A法)"进行配对比较实验。采用A,B法分别采集同一只小鼠的双侧附睾精子。结果前向运动精子活跃地从"双舱培养皿"的内舱游出至外舱。A、B两法分离得到的前向运动精子百分率分别为(56.83±7.23)%和(77.33±5.69)%,变异系数分别为12.72%和7.36%,差别具有统计学意义。结论"B法"能稳定显著地提高前向运动精子的百分率,优于"A法"。

Objective The technique of mouse sperm collection is common used in many fields of life science currently. The aim of this research is in order to improve the ratio of forward motile sperm collecting from mouse epididymides. Method A novel ＂double chambers dish＂ and convenient ＂double chambers swim＂ method were designed and compared with ＂conventional swim-up＂ method which is in common used nowadays. Experiments showed that the forward motile sperm swim-out actively from the inner-chamber and the rate of forward motile sperm increased. Sperm was collected from different side of the same mouse by using ＂conventional swim-up＂ or ＂double chambers swim＂ method,respectively. Result The means of the rate of forward motile sperm collected were 56.83%-4-7, 23% with CV12. 72% in ＂conventional swim-up＂ group and 77.33%-4-5.69% with CVT. 36% in ＂double chambers swim＂ group, respectively. Statistical analysis showed there are significant differences between two datas according to the paired t-test. Conclusion The ＂double chambers swim＂ method can improve the ratio of the forward motile sperm stably and observably and would be a better method in mouse sperm collection.