Abstract

Abstract Study question Does HyperSperm improve sperm capacitation without affecting sperm quality? Summary answer HyperSperm enhances sperm hyperactivated motility, a hallmark of capacitation, without compromising sperm viability, acrosome integrity, or DNA fragmentation. What is known already HyperSperm is a novel semen preparation method for IVF/ICSI based on mimicking effective capacitation that occurs in the female tract. HyperSperm, which consists of 3 media used sequentially to treat sperm before insemination, gave rise to more high-quality blastocysts compared to current standard of care in a pilot split oocyte study in 10 couples. Study design, size, duration This prospective in vitro study was conducted between July 2023 and January 2024 in an academic department. A consecutive cohort of 15 normozoospermic individuals were recruited and included. Each participant provided a semen sample by masturbation after written informed consent was obtained on the day of the collection. Participants/materials, setting, methods Semen samples from normozoospermic individuals (n = 15) were divided into two halves. After density gradient centrifugation, Control half was incubated in modified HTF (Human Tubal Fluid) for 2-4 hours, while the other half underwent HyperSperm treatment. The following parameters were assessed: sperm viability (eosin exclusion); motility and kinematic parameters (CASA); acrosome integrity (fluorescein-labeled Pisum sativum lectin staining); and DNA fragmentation by TUNEL. Differences between groups were evaluated by paired T-test. Significant differences were considered when p < 0.05. Main results and the role of chance Following treatments, sperm viability (90±3% Control vs. 91±3% HyperSperm) and motility (87±2% Control vs. 89±3% HyperSperm) were similar between groups (p > 0.05). Notably, HyperSperm induced a significant hyperactivation increase (9±2% Control vs. 22±3% HyperSperm), along with associated kinematic parameters (i.e., curvilinear velocity and lateral head displacement) (p < 0.05). Furthermore, HyperSperm treatment did not promote motility decrease over time. Finally, there were no differences in the percentage of sperm with intact acrosome (91±2% Control vs. 90±2% HyperSperm) and DNA fragmentation (3±1% Control vs. 3±1% HyperSperm) between groups (p > 0.05). Altogether, HyperSperm has shown to significantly improve sperm capacitation parameters while maintaining a high safety profile in the treated sample. Limitations, reasons for caution The study is limited to normozoospermic samples, and further research should validate these findings across broader male populations undergoing IVF/ICSI. Wider implications of the findings HyperSperm has demonstrated an increase in the production of high-quality blastocysts via IVF, that would reduce barriers to treatment access through higher cumulative success rates and lower treatment repetitions. The observed increase in hyperactivation combined with the safety profile of HyperSperm supports its potential for future adoption in reproductive clinics. Trial registration number not applicable

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