Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.
Hum Reprod. 2010 Apr;25(4):839-46. doi: 10.1093/humrep/deq009. Epub 2010 Feb 10.
Couples with unexplained infertility (UI) tend to have low fertilization rates with current IVF procedures. Here, we attempted to identify spermatozoa with apoptotic markers in couples with UI and unsuccessful intrauterine insemination (IUI) and we investigated the efficiency and benefit of magnetic-activated cell sorting (MACS) for sperm preparation in such patients.
Sixty couples with UI and two IUI failures were recruited. The sperm were prepared by conventional density gradient centrifugation (DGC) and divided into two aliquots. One aliquot was used as a control and the other was further processed by MACS (D + M). Apoptotic markers were identified using fluorescence-labeled dye and flow cytometry, including externalization of phosphatidylserine (EPS), disrupted mitochondrial membrane potential (MMP) and DNA fragmentation. The fertilization potential of prepared spermatozoa was analyzed by basic semen analysis, computer-aided sperm analysis and the induced acrosome reaction test (IART).
After DGC, spermatozoa showed 18.6% EPS, 28.3% disrupted MMP and 13.5% DNA fragmentation. Numbers of spermatozoa with apoptotic markers were significantly reduced by D + M, versus DGC alone (P < 0.001). Although the motility of spermatozoa was slightly decreased after MACS, most sperm motion characteristics were not impaired. Interestingly, the IART significantly improved after D + M, versus DGC alone, especially for the couples with a normal hemizona assay (P < 0.001).
The spermatozoa prepared by D + M showed a reduced level of apoptotic markers. Improvement in the IART suggests a high fertilization potential of the processed spermatozoa. The identification of apoptotic markers and use of MACS may be helpful in directing the management plan for patients with UI and multiple IUI failures.
不明原因不孕（UI）的夫妇在当前的试管婴儿（IVF）程序中受精率往往较低。在这里，我们试图在 UI 和两次宫腔内人工授精（IUI）失败的夫妇中识别具有凋亡标记的精子，并研究磁激活细胞分选（MACS）在这些患者的精子准备中的效率和益处。
招募了 60 对 UI 和两次 IUI 失败的夫妇。通过常规密度梯度离心（DGC）和分成两份来制备精子。一份作为对照，另一份用 MACS（D+M）进一步处理。通过荧光标记染料和流式细胞术鉴定凋亡标记物，包括磷脂酰丝氨酸（EPS）的外翻、线粒体膜电位（MMP）的破坏和 DNA 片段化。通过基础精液分析、计算机辅助精子分析和诱导顶体反应试验（IART）分析制备精子的受精潜力。
DGC 后，精子显示 18.6%的 EPS、28.3%的破坏 MMP 和 13.5%的 DNA 片段化。与 DGC 相比，D+M 显著减少了具有凋亡标记的精子数量（P<0.001）。尽管 MACS 后精子的活力略有下降，但大多数精子运动特征并未受损。有趣的是，与 DGC 相比，IART 显著改善，特别是对于正常半透明带检测的夫妇（P<0.001）。
D+M 制备的精子显示出较低水平的凋亡标记。IART 的改善表明处理后的精子具有较高的受精潜力。凋亡标记的鉴定和 MACS 的使用可能有助于指导 UI 和多次 IUI 失败患者的管理计划。
