Kim Sung Woo, Jee Byung Chul, Kim Seul Ki, Kim Seok Hyun
Department of Obstetrics and Gynecology, Seoul National University Hospital, Seoul, Korea.
Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea.
Clin Exp Reprod Med. 2017 Dec;44(4):201-206. doi: 10.5653/cerm.2017.44.4.201. Epub 2017 Dec 31.
The aim of this study was to compare the efficacy of swim-up and density gradient centrifugation (DGC) for reducing the amount of sperm with fragmented DNA, sex chromosome aneuploidy, and abnormal chromatin structure.
Semen samples were obtained from 18 healthy male partners who attended infertility clinics for infertility investigations and were processed with swim-up and DGC. The percentages of sperm cells with fragmented DNA measured by the sperm chromatin dispersion test, normal sex chromosomes assessed by fluorescence hybridization, and abnormal chromatin structure identified by toluidine blue staining were examined.
The percentage of sperm cells with fragmented DNA was significantly lower in the swim-up fraction (9.7%, =0.001) than in the unprocessed fraction (27.0%), but not in the DGC fraction (27.8%, =0.098). The percentage of sperm cells with normal X or Y chromosomes was comparable in the three fractions. The percentage of sperm cells with abnormal chromatin structure significantly decreased after DGC (from 15.7% to 10.3%, =0.002). The swim-up method also tended to reduce the percentage of sperm cells with abnormal chromatin structure, but the difference was not significant (from 15.7% to 11.6%, =0.316).
The swim-up method is superior for enriching genetically competent sperm.
本研究旨在比较上游法和密度梯度离心法(DGC)在减少DNA碎片化、性染色体非整倍体和染色质结构异常精子数量方面的效果。
从18名因不育问题前往不孕不育门诊就诊的健康男性伴侣处获取精液样本,分别采用上游法和DGC法进行处理。检测通过精子染色质扩散试验测定的DNA碎片化精子细胞百分比、通过荧光杂交评估的正常性染色体以及通过甲苯胺蓝染色鉴定的染色质结构异常情况。
上游法处理后的精子样本中,DNA碎片化精子细胞的百分比(9.7%,P = 0.001)显著低于未处理样本(27.0%),但DGC法处理后的样本(27.8%,P = 0.098)与未处理样本相比无显著差异。三个样本中具有正常X或Y染色体的精子细胞百分比相当。DGC法处理后,染色质结构异常的精子细胞百分比显著降低(从15.7%降至10.3%,P = 0.002)。上游法也倾向于降低染色质结构异常的精子细胞百分比,但差异不显著(从15.7%降至11.6%,P = 0.316)。
上游法在富集具有遗传活性的精子方面更具优势。