Zanjan Metabolic Diseases Research Center, Zanjan University of Medical Sciences, Zanjan, Iran.
Department of Obstetrics and Gynecology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Biopreserv Biobank. 2024 Feb;22(1):38-45. doi: 10.1089/bio.2022.0174. Epub 2023 Oct 6.
The cryopreservation-thawing process of spermatozoa cells has negative impacts on their structure, function, and fertility parameters, which are known as cryoinjury. Asthenozoospermia patients are more susceptible to cryoinjury. Granulocyte-macrophage colony-stimulating factor (GM-CSF) increases sperm glucose uptake via the induction of glucose transporters, resulting in increased sperm motility. This study aimed to investigate the efficiency of GM-CSF supplementation of the cryopreservation media for semen samples of asthenoteratozoospermia patients. The study was carried out on 20 semen samples from infertile men referred to diagnosing semen analysis. To avoid subjective bias, two main sperm motility parameters, including velocity along the curvilinear path and velocity along the straight-line path were considered by the computer-assisted sperm analysis system. Afterward, each semen sample was divided into three equal aliquots and randomly assigned to one of the following groups: group I (control, freezing media only), group II (+GM-CSF, freezing medium supplemented with 2 μL/mL GM-CSF), or group III (GM-CSF added after thawing and washing). Following semen thawing, standard parameters, mitochondrial membrane potential (MMP), and the DNA Fragmentation Index were analyzed. Total sperm motility (progressive and non-progressive) improved significantly in group III samples after a 30-minute incubation with GM-CSF compared with the control group (26.5% ± 3.1% vs. 17.51% ± 2.59%). However, no differences in progressive motility or sperm morphology were found among the three thawed samples. The percentage of vitality was significantly higher in group III compared with the other two groups (28.38% ± 3.4% vs. 22.4% ± 3.08% and 22.14% ± 2.77%, respectively) ( < 0.05). JC-1 levels (a marker of MMP) were not significantly different between the examined groups (44.95% ± 8.26% vs. 36.61% ± 6.95% vs. 46.67% ± 7.7%, for control, group II, and group III, respectively) ( > 0.05). GM-CSF may be advantageous as an additive after freezing, improving total motility and viability after 30 minutes of post-thaw incubation; however, when supplied to the freezing media before cryopreservation, it is unable to protect against cryoinjury.
精子细胞的冷冻-解冻过程对其结构、功能和生育参数有负面影响，这被称为冷冻损伤。弱精症患者更容易受到冷冻损伤的影响。粒细胞-巨噬细胞集落刺激因子（GM-CSF）通过诱导葡萄糖转运体增加精子的葡萄糖摄取，从而提高精子的运动能力。本研究旨在探讨 GM-CSF 对弱精症患者精液样本冷冻保存液的补充效率。该研究共纳入 20 例不育男性的精液样本，用于精液分析诊断。为了避免主观偏见，计算机辅助精子分析系统考虑了两个主要的精子运动参数，即曲线运动速度和直线运动速度。之后，将每个精液样本均分为 3 等份，并随机分配到以下 3 个组之一：组 I（对照组，仅冷冻保存液）、组 II（+GM-CSF，冷冻保存液中添加 2μL/mL GM-CSF）或组 III（解冻和洗涤后添加 GM-CSF）。解冻后，分析标准参数、线粒体膜电位（MMP）和 DNA 片段化指数。与对照组相比，解冻后 30 分钟孵育 GM-CSF 的组 III 样本中的总精子运动（前向和非前向）显著提高（26.5%±3.1%对 17.51%±2.59%）。然而，三组解冻样本的前向运动或精子形态无差异。与其他两组相比，组 III 的活力百分比明显更高（28.38%±3.4%对 22.4%±3.08%和 22.14%±2.77%）（ <0.05）。实验组间 JC-1 水平（MMP 的标志物）无显著差异（44.95%±8.26%对 36.61%±6.95%对 46.67%±7.7%，分别为对照组、组 II 和组 III）（ >0.05）。GM-CSF 作为冷冻后的添加剂可能是有益的，可在解冻后 30 分钟的孵育后提高总运动能力和活力；然而，在冷冻保存前添加到冷冻保存液中时，它不能防止冷冻损伤。
