Al Hadi Laboratory and Medical Centre, Beirut, Lebanon.
Holy Spirit University of Kaslik (USEK), Jounieh, Lebanon.
Andrology. 2021 Sep;9(5):1549-1559. doi: 10.1111/andr.13038. Epub 2021 Jun 18.
Suboptimal human semen handling in vitro may induce sperm damage. However, the effects of semen swim-up, pellet swim-up, density gradient, and density gradient followed by SU on sperm motility, morphology, DNA fragmentation, acrosome reaction, intracellular reactive oxygen species, and mitochondrial activity were not fully understood.
To study the impact of four sperm preparation techniques on sperm functional parameters.
This study was conducted on 60 infertile men with a minimum sperm concentration of 20 × 10 /ml and total sperm motility of ≥30%. Each raw semen sample was divided into four aliquots. Each aliquot was prepared by one of the tested techniques. Various sperm characteristics were assessed before and after sperm preparation.
Density gradient and density gradient followed by SU resulted in significantly higher DNA fragmentation percentages compared with semen swim-up (p < 0.001 and p < 0.001, respectively) and pellet swim-up (p < 0.001 and p < 0.001, respectively). Significantly higher percentages of spermatozoa with intact acrosome were detected in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen. The percentage of reactive oxygen species-positive spermatozoa was significantly higher after pellet swim-up (p < 0.001), density gradient (p < 0.001), and density gradient followed by SU (p < 0.001) than raw semen. In addition, the percentages of 100% stained midpiece (active mitochondria) were significantly higher in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen.
To the best of our knowledge, this is the first report comparing the impact of these techniques on various sperm functional parameters. Semen swim-up was more effective than density gradient in selecting better spermatozoa in terms of DNA integrity, reactive oxygen species levels, acrosome status, and mitochondrial activity. Randomized clinical trials comparing these four techniques are required to test their impact on embryo development and pregnancy outcomes.
体外处理不佳的人类精液可能会导致精子损伤。然而,精液游移、沉淀游移、密度梯度和密度梯度后游移对精子活力、形态、DNA 碎片化、顶体反应、细胞内活性氧和线粒体活性的影响尚未完全了解。
研究四种精子制备技术对精子功能参数的影响。
本研究纳入了 60 名最低精子浓度为 20×10/ml 且总精子活力≥30%的不育男性。每个原始精液样本被分为四份。每份样本通过一种测试技术进行处理。在精子处理前后评估各种精子特征。
与精液游移(p<0.001 和 p<0.001)和沉淀游移(p<0.001 和 p<0.001)相比,密度梯度和密度梯度后游移导致 DNA 碎片化百分比显著增加。与原始精液相比,在精液游移(p<0.001)和沉淀游移(p<0.001)中,具有完整顶体的精子百分比显著增加。与原始精液相比,经过沉淀游移(p<0.001)、密度梯度(p<0.001)和密度梯度后游移(p<0.001)后,活性氧阳性精子的百分比显著增加。此外,在精液游移(p<0.001)和沉淀游移(p<0.001)中,100%染色中段(活跃线粒体)的百分比显著高于原始精液。
据我们所知,这是第一项比较这些技术对各种精子功能参数影响的报告。与密度梯度相比,精液游移在 DNA 完整性、活性氧水平、顶体状态和线粒体活性方面更有效地选择了更好的精子。需要进行随机临床试验来比较这些技术对胚胎发育和妊娠结局的影响。