This record contains raw data related to article “New methodology based on freeze-drying for long-term preservation of human sperm" Freeze-drying is an ecological non-cryogenic storage solution that could represent an economic tool to transport sperm all over the world. This methodology was applied only to mice and a few other species so far, but not for a long-term storage of human sperm. For the first time we investigated whether freeze-drying could deteriorate human sperm DNA fragmentation at different time points (1 month, 3 months and 10 months). We used a high concentration of trehalose in the freeze-drying buffer and 2 vials per time point were insufflated with argon, replacing oxygen, and stored at 4°C or room temperature; the remaining 2 vials were stored in normal atmosphere at the same temperature conditions. We did not observe any statistically significant difference between the groups analyzed, suggesting that our lyophilization solution could maintain unaltered sperm DNA fragmentation until 10 months of storage without the presence of a high vacuum. These evidences support the potential use of freeze-drying also for human sperm in fertility clinics. Correspondence: Stefano Castellano, Division of Gynecology and Reproductive Medicine, Department of Gynecology, Fertility Center, Humanitas Research Hospital – IRCCS, Via Alessandro Manzoni 56, 20089, Rozzano, Milan, Italy (mobile: +393347401507, email: stefano.castellano@humanitas.it)