Comparative Efficacy of Swim-Up, Density-Gradient Centrifugation, and Microfluidic Sorting in Sperm Preparation, and the Impact on Motility, Morphology, and DNA Integrity

作者：Zina Wen, Li Duan, Yong Chen, Qing-Hong Qiu, Gang Liu, Ning Luo, Peng-Hao Li, Er-Po Tian, Ren‐Shan Ge · 发表于：International Journal of General Medicine · 年份：2025 · DOI：10.2147/ijgm.s517575 · 被引用次数：7 · 研究领域：Sperm and Testicular Function、Reproductive Biology and Fertility、Reproductive Health and Technologies

Background: Reactive oxygen species (ROS) are considered a major factor contributing to sperm DNA damage during sperm preparation for assisted reproductive technologies (ART). This study aimed to investigate whether microfluidic sorting can select sperm with a low DNA fragmentation index (DFI) and to explore the underlying mechanisms. We compared the effects of three sperm preparation methods—swim-up, density-gradient centrifugation, and microfluidic sorting—on sperm quality and DNA integrity. Methods: Semen samples from 12 patients were divided into three equal portions and processed using swim-up, density-gradient centrifugation, and microfluidic sorting techniques. Sperm concentration, motility, morphology, DFI, intracellular H 2 O 2 levels, and mitochondrial O 2 ⁻ levels were measured and compared across the three methods. Additionally, DFI was assessed in both fresh and frozen-thawed sperm samples. Results: Sperm prepared using microfluidic sorting exhibited significantly higher total motility (85.3 ± 3.2%) and progressive forward motility (72.5 ± 2.8%) compared to density-gradient centrifugation (total motility: 70.1 ± 3.5%; progressive motility: 58.4 ± 3.1%). Microfluidic sorting also resulted in a significantly lower DFI (8.2 ± 1.5%) compared to density-gradient centrifugation (25.6 ± 2.3%) and swim-up (15.4 ± 1.8%). Intracellular H 2 O 2 levels were similar across all methods, but mitochondrial O 2 ⁻ levels were significantly lower in microfluidic-sorted sperm (12.3 ...