Rosenborg L, Rao K M, Björndahl L, Kvist U, Pousette A, Akerlöf E, Fredricsson B
Department of Obstetrics and Gynecology, Huddinge University Hospital, Sweden.
Int J Androl. 1990 Aug;13(4):287-96. doi: 10.1111/j.1365-2605.1990.tb01034.x.
This study was designed to define the effects of sperm preparation on sperm chromatin stability in relation to in-vitro fertilization (IVF). Semen samples used for IVF-embryo transfer (ET) in the treatment of infertility due to tubal factors were studied. Cases with semen variables below reference limits in previous samples were excluded. Sperm were prepared by a swim-up technique employing either of two different tissue culture media, Ham's F-10 or Earle's balanced salt solution. Sperm chromatin stability was tested by exposure both to sodium dodecyl sulphate (SDS) only and SDS together with a zinc-chelating agent, disodium ethylene diamine tetraacetate (SDS-EDTA). Sperm head swell scores were defined under different experimental conditions and the relationship to sperm motility, morphology, fertilization rate and pregnancy occurrence was tested. No differences were seen between the chromatin stability of sperm from the original sample and that after swim-up preparation, neither immediately after completion of the swim-up procedure, nor at the time of insemination of ova. With time, the chromatin became more stable, which occurred to a similar extent both in the original sample and in swim-up preparations using Ham's F-10. Otherwise, sperm chromatin stability was unaffected by either of the two media used for swim-up. At higher incubation temperatures, decondensation in SDS was enhanced. Altogether, no correlation was found between sperm chromatin stability or enhancement of decondensation by temperature and the success of IVF treatment expressed in fertilization rates or pregnancies. The results are reassuring in that only small changes in sperm chromatin stability occurred during the preparation for IVF. As long as semen of presumably good quality is used, these changes in chromatin stability do not seem to be of clinical importance.
本研究旨在确定精子制备对与体外受精(IVF)相关的精子染色质稳定性的影响。对因输卵管因素导致不孕而用于IVF-胚胎移植(ET)的精液样本进行了研究。排除先前样本中精液变量低于参考限值的病例。采用上浮技术,使用两种不同的组织培养基(Ham's F-10或Earle平衡盐溶液)之一制备精子。通过仅暴露于十二烷基硫酸钠(SDS)以及SDS与锌螯合剂乙二胺四乙酸二钠(SDS-EDTA)共同作用来测试精子染色质稳定性。在不同实验条件下定义精子头部肿胀评分,并测试其与精子活力、形态、受精率和妊娠发生率的关系。在原样本精子与上浮制备后的精子染色质稳定性之间,无论是在上浮程序完成后立即观察,还是在卵子授精时观察,均未发现差异。随着时间推移,染色质变得更加稳定,在原样本以及使用Ham's F-10进行上浮制备的样本中,这种情况的发生程度相似。此外,用于上浮的两种培养基中的任何一种均未影响精子染色质稳定性。在较高的孵育温度下,SDS中的去浓缩作用增强。总体而言,未发现精子染色质稳定性或温度引起的去浓缩增强与以受精率或妊娠表示的IVF治疗成功率之间存在相关性。这些结果令人安心,因为在IVF准备过程中精子染色质稳定性仅发生了微小变化。只要使用质量大概良好的精液,染色质稳定性的这些变化似乎并无临床重要性。