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In recent years, there has been an increased interest in new preservation techniques that facilitate sperm storage and distribution, with freeze-drying (FD) having been proposed as an alternative method for sperm preservation and maintenance of genetic resources in different animal species. FD is a method in which frozen material is dried by sublimation of ice, thereby involving a direct transition from a solid (ice) to a vapour (gas) phase. One of the main advantages of FD is that nitrogen and dry ice are no longer required for the storage and shipment of frozen sperm, which can be stored at room temperature or 4°C, thereby resulting in enormous reductions in storage and shipping costs. Unlike sperm cryopreserved after gradual freezing, the sperm membrane may be further damaged by both snap-freezing and drying stresses during the FD procedure. As mammalian spermatozoa lose their motility, viability and, at least partially, their DNA integrity when freeze-dried, they must be microinjected into an oocyte by intracytoplasmic sperm injection (ICSI). Although the efficiency of ICSI is limited when freeze-dried spermatozoa are used, embryos and live offspring can be produced. DNA fragmentation in freeze-dried spermatozoa is one of the main causes of failure of embryonic development and successful pregnancy. In this regard, it has been suggested that endonucleases are among the leading causes of DNA fragmentation in spermatozoa along with oxidative stress caused by the release of reactive oxygen species (ROS). Many factors influence the FD process, and it is not clear how FD affects specific components of sperm from different animal species. As such, a sound understanding of the FD process would result in increased production of embryos and/or live offspring. The aim of this review was to study the various stages and techniques used in the FD process and to further evaluate the results obtained.