Microwave-Assisted Dehydration, Non-Cryogenic Storage, and Rehydration of Spermatozoa and Germinal Vesicles

The microwave-assisted dehydration technique enables preservation of both male and female genomes in a dried state at non-cryogenic temperatures, eliminating the reliance on liquid nitrogen for long-term storage. The dehydration strategy outlined below utilizes trehalose, a natural disaccharide known to ensure survival of certain primitive species under drought conditions, to increase the dehydration tolerance of the cells. In this chapter, we describe step-by-step procedures for trehalose loading, microwave-assisted dehydration, sample storage, and rehydration of both spermatozoa and germinal vesicle stage oocytes. Tips for modification are provided to apply the protocols to different species of interest.