生殖医学杂志

目的探讨洗涤法(SW)、密度梯度离心法(DGC)、上游法(SU)、磁性活性细胞分选法(MACS)对冷冻精液的优选效果,为供精人工授精(AID)和供精体外受精(IVF-D)提供可靠的数据。方法门诊收集健康男性精液,制备冷冻精液。冷冻精液复苏后,根据优选处理方法不同分为4组:SW组、DGC组、SU组和MACS组,每组样本各20例。比较分析各组优选前后精子浓度、前向运动精子活率、膜完整性、正常形态率及DNA碎片率等的差异。结果 SW组,处理前后前向运动精子活率、精子膜完整性、正常形态率及DNA完整率,均无显著性差异(P>0.05);DGC和SU组中,优选后前向运动精子活率[分别为(78.19±12.13)vs.(34.17±8.61)、(82.20±7.17)vs.(42.86±16.38)]、精子膜完整率[分别为(84.62±5.84)vs.(68.17±6.74)、(81.44±4.47)vs.(65.19±6.15)]、正常形态率[(20.42±4.63)vs.(12.16±5.75)、(25.02±4.61)vs.(10.24±5.37)]及DNA完整率[(86.73±7.07)vs.(67.98±6.45)、(88.49±3.09)vs.(67.44±5.89)]均显著高于同组优选前(P<0.05);MACS组中,优选后冷冻精子的膜完整性[(84.00±5.33)vs.(69.21±7.79)]、正常形态率[(13.72±4.58)vs.(7.90±3.80)]及DNA完整率[(91.95±2.82)vs.(69.79±3.90)]亦显著高于优选前(P<0.05),但前向运动精子活率没有显著改善(P>0.05)。结论四种方法在处理冷冻精子方面各有优缺点,临床可以根据精液标本的质量或者对于精液制备液的要求来选择不同的精子优化方法。MACS可以较好地优化DNA完整率,其在辅助生殖领域的应用具有一定前景。

Objective:To compare the effect of optimizing frozen-thawed sperm by using simple washing(SW),swim-up(SU),density-gradient centrifugation(DGC)and magnetic activated cell sorting(MACS)for providing reliable data for AID and IVF-D.Methods:The semen samples of healthy men were collected and frozen.The frozen-thawed semen samples were divided into 4 groups:SW,DGC,SU and MACS group according to the optimizing method,20 samples for each group.The concentration,motility,membrane integrity,normal morphology rate and DNA fragment rate were compared and analyzed among the four groups.Results:There was no significant differences in motility,membrane integrity,normal morphology rate and DNA fragment rate before and after treatment as using SW method(P>0.05).In DGC and SU groups,the membrane integrity rate[(84.62±5.84)vs.(68.17±6.74)and(81.44±4.47)vs.(65.19±6.15),respectively],normal morphology rate[(20.42±4.63)vs.(12.16±5.75),(25.02±4.61)vs.(10.24±5.37)]and DNA integrity rate[(86.73±7.07)vs.(67.98±6.45),(88.49±3.09)vs.(67.44±5.89)]after treatment were significantly higher than those before treatment(P<0.05).In MACS method group,the membrane integrity rate[(84.00±5.33)vs.(69.21±7.79)],normal morphology rate[(13.72±4.58)vs.(7.90±3.80)]and DNA integrity rate[(91.95±2.82)vs.(69.79±3.90)]after treatment were significantly higher than those before treatment(P<0.05),but there was no significant improvement in sperm progressive motility(P>0.05).Conclusions:The four methods have their own advantage and disadvantage in treatment of frozenthawed sperm.The sperm optimization method should be selected according to the quality of semen or semen preparation requirement.MACS can optimize DNA integrity rate and its application in the field of assisted reproduction will has certain prospects.