Abstract：
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) (p < 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) (p > 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.
摘要：
精子细胞的冷冻保存-解冻过程对其结构有负面影响,函数,和生育率参数，这被称为冷冻损伤。无精症患者更容易发生冷冻损伤。粒细胞-巨噬细胞集落刺激因子（GM-CSF）通过诱导葡萄糖转运蛋白增加精子葡萄糖摄取，导致精子活力增加。本研究旨在研究补充GM-CSF冷冻保存培养基对弱精子症患者精液样本的效率。这项研究是针对20例不育男性的精液样本进行的，这些样本涉及诊断精液分析。为了避免主观偏见，两个主要的精子运动参数，计算机辅助精子分析系统考虑了沿曲线路径的速度和沿直线路径的速度。之后,将每个精液样本分为三个相等的等分试样，并随机分配到以下一组：I组（对照，仅限冷冻介质)，II组（+GM-CSF，补充有2μL/mLGM-CSF的冷冻培养基），或III组（解冻和洗涤后添加GM-CSF）。精液解冻后,标准参数,线粒体膜电位（MMP），分析DNA片段化指数。与对照组相比，与GM-CSF孵育30分钟后，第III组样品的总精子运动性（进行性和非进行性）显着提高（26.5％±3.1％vs.17.51%±2.59%）。然而,在三个解冻的样本中，没有发现进行性运动或精子形态的差异。与其他两组相比，III组的活力百分比明显更高（28.38％±3.4％vs.22.4%±3.08%和22.14%±2.77%，分别)(p<0.05)。JC-1水平（MMP的标志物）在检查组之间没有显着差异（44.95％±8.26％vs.36.61%±6.95%vs.46.67%±7.7%，为了控制，第二组,第三组,分别)(p>0.05)。GM-CSF作为冷冻后的添加剂可能是有利的，解冻后孵育30分钟后，改善总运动性和生存力；然而，当在冷冻保存之前提供给冷冻介质时，它无法防止冷冻损伤。
