1层和2层梯度离心法分离精子的效果评价

Single-and two-layer gradient centrifugation in sperm separation:Comparison and appraisal

目的:评价1层和2层Percoll密度梯度离心法分离精子的效果。方法:20份精液标本分别行50%1层,90%和45%Percoll2层密度梯度离心分离,处理前后应用SCA(sperm class analyzer)精子质量分析仪分析精子密度、活力和圆形细胞密度。结果:1层法分离后精子回收率为(65.5±12.8)%,明显高于2层法(P<0.01);1层和2层法分离后a级精子百分率明显高于处理前(P<0.05,P<0.01),而1层法分离后a级精子百分率明显低于2层法(P<0.05);1层法分离精子后c级精子百分率明显高于2层法(P<0.05),与处理前相比没有明显差异(P>0.05);2层法分离后a+b级精子百分率明显高于处理前(P<0.05),1层法分离后a+b级精子百分率与处理前相比没有明显差异(P>0.05);1层和2层法分离后圆形细胞密度明显低于处理前(P<0.05,P<0.01),两种方法之间没有差异(P>0.05)。结论:1层法分离后精子回收率较高,精子的活力改变不大;2层法分离后精子回收率较低,精子的活力明显改善;1层和2层法都可以较好地把精子与圆形细胞分开。两种方法各有优势,在精子体外处理中都有着重要的应用价值。

Objective：To appraise the effect of single-and two-layer Percoll density gradient centrifugation in sperm separation.Methods：Twenty semen specimens underwent single-（50%）and two-layer（90% and 45%）density gradient centrifugation,respectively.The sperm class analyzer（SCA）was used to analyze sperm density,motility and dynamic parameters and round cell density before and after the treatment.Results：After separation,the sperm recovery rate of the single-layer method was（65.5±12.8）%,significantly higher than that of the two-layer method（P0.01）.The percentages of grade a sperm of the single-and two-layer method were significantly higher than pre-treatment（P0.05,P0.01）,that of the single-layer was significantly lower than that of the two-layer method（P0.05）,but the percentage of grade c sperm of the former was significantly higher than that of the latter（P0.05）.Compared with pre-treatment,the percentage of grade a＋b sperm of the two-layer method was significantly higher（P0.05）,while that of the single-layer method showed no significant difference（P0.05）,and the round cell density of both the methods was significantly lower（P0.05,P0.01）,with no significant differences between the two methods（P0.05）.Conclusion：The single-layer method yields a higher rate of sperm recovery and causes little change in the sperm motility,while the two-layer method effects a lower rate and significantly improves sperm motility.Both the methods can efficiently separate sperm from round cells,and each has its own advantages and its application value in in vitro treatment of sperm.