The freeze-drying of sperm has been developed as a new preservation method where liquid nitrogen is no longer necessary. An advantage of freeze-drying sperm is that it can be stored at 4°C and transported at room temperature. We showed that the fertility of freeze-dried sperm could be maintained for 5 years without deterioration. Offspring with normal fertility were generated from oocytes fertilized with sperm freeze-dried in Tris-EDTA buffer. This is the first report to demonstrate the successful freeze-drying of sperm using this new and simple method for long-term preservation. Furthermore, freeze-dried samples can be temporarily stored at room temperature even in the event of a power failure, interruption to the liquid nitrogen supply or other emergencies caused by disasters such as earthquakes and typhoons. We strongly believe that the freeze-drying process provides a safe and economical preservation of valuable animal strains, and provides us with a new method of sperm preservation for bio-banking.