透明带选择后的精子形态和 DNA 片段。

Reproduction & Fertility Pub Date : 2021-08-12 eCollection Date: 2021-07-01 DOI:10.1530/RAF-21-0041
Rumiana Ganeva, Dimitar Parvanov, Denitsa Velikova, Magdalena Vasileva, Kristina Nikolova, Georgi Stamenov
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引用次数: 0

摘要

精子 DNA 片段(SDF)和精子形态缺陷会对 ART 的结果产生负面影响。因此,在进行卵胞浆内单精子显微注射(ICSI)前,需要额外的精液处理技术,以考虑精子的DNA状态和形态。该研究的目的是评估基于透明带粘附的精子选择技术与原生精液和常规游动精子制备技术相比,在获得具有高比例正常形态和DNA完整性的精子群体方面的功效。对 78 名正常无精子男性的精液样本进行上浮处理,并将其置于涂有 48 种酸溶解的透明带的培养皿中。对每位患者的原生精液、上浮精液样本和涂有透明带的精子的精子 DNA 片段和形态进行了评估。分区精子的平均精子DNA片段率(3.5 ± 0.7%)明显低于游动样本(15.3 ± 5.2%)(P < 0.001)和本地精液(24.9 ± 7.1%)(P < 0.001)。经过透明带粘附的额外筛选后,所有样本的DNA损伤水平都较低。与游动样本(8.9 ± 4.3%)(P < 0.001)或原生精液(5.3 ± 3.2%)(P < 0.001)相比,经过透明带粘附选择后,形态正常的精子比例明显更高(11.4 ± 3.9%)。在94%的样本中,经过额外的透明带选择后,具有正常形态的精子比例有所增加。该研究表明,与原生精液和纯游泳制备的样本相比,通过附加透明带粘附技术进行精子选择后,具有正常形态的精子比例明显提高,DNA碎片水平也明显降低。因此,有必要在辅助生殖治疗前采用更多的精液处理技术来检测精子质量。我们的团队设计了一种简单的技术,利用卵子外膜(透明带)中的蛋白质,在精子与卵子自然相互作用的基础上加强精子选择程序。将这项技术与人工生殖技术中最常用的技术结合使用,可显著提高具有正常形状、大小和结构的精子的比例,并减少 DNA 片段的数量。如果将这种精子区域选择技术引入人工授精实践,将有利于获得受精潜能更高的精子。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Sperm morphology and DNA fragmentation after zona pellucida selection.

Sperm morphology and DNA fragmentation after zona pellucida selection.

Sperm morphology and DNA fragmentation after zona pellucida selection.

Sperm morphology and DNA fragmentation after zona pellucida selection.

Sperm DNA fragmentation (SDF) and sperm morphological defects can negatively affect ART outcomes. Consequently, there is a need for additional semen processing technique that accounts for sperm DNA status and morphology prior to ICSI. The objective was to evaluate the efficacy of an additional zona pellucida adhesion-based sperm selection for obtaining sperm populations with a high percentage of normal morphology and DNA integrity as compared to native semen and routine swim-up preparation. Semen samples from 78 normozoospermic men were subjected to swim up and placed in petri dishes coated with 48 acid-solubilized zonae pellucidae. Sperm DNA fragmentation and morphology were assessed in the native semen, the swim-up samples, and the zona-adhered spermatozoa from each patient. The mean sperm DNA fragmentation of the zona-selected spermatozoa (3.5 ± 0.7%) was significantly lower than the swim-up samples (15.3 ± 5.2%) (P  < 0.001) and native semen (24.9 ± 7.1%) (P  < 0.001). All of the samples had lower levels of DNA damage after additional selection by zona pellucida adhesion. Significantly higher percentage of sperm with normal morphology was observed after zona-adhesion selection (11.4 ± 3.9%) when compared to the swim-up samples (8.9 ± 4.3%) (P  < 0.001) or the native semen (5.3 ± 3.2%) (P  < 0.001). In 94% of the samples, the percentage of spermatozoa with normal morphology increased after the additional zona selection. This study demonstrates that sperm selection by additional zona-adhesion technique yields a significantly higher percentage of spermatozoa with normal morphology as well as a significantly decreased level of DNA fragmentation when compared to the native semen and the swim-up-only prepared samples.

Lay summary: High level of DNA folding known as sperm DNA fragmentation (SDF) inside each sperm and defects in the shape, size, and structure of the sperm can negatively affect assisted reproduction treatment (ART) outcomes. Consequently, there is a need for additional semen processing techniques that account for sperm quality prior to ART. Our team designed a simple technique using proteins from the coat around the egg (zona pellucida) to enhance sperm selection procedures based on natural sperm-egg interactions. Using this technique in combination with the most common techniques used in ART yields a significantly higher percentage of sperm with normal shape, size, and structure and a decreased level of DNA fragmentation. This sperm zona-selection technique would be beneficial if introduced in the ART practice to yield sperm with higher fertilization potential.

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