Sperm preservation is an important technique for maintaining valuable genetic resources in biomedical research and wildlife. In the mouse, the sperm cryopreservation method has been established and adopted by large-scale sperm preservation projects in cryobanks. Recently, a new sperm preservation method using freeze-drying has been studied in various mammals. Freeze-drying is the ultimate method by which sperm can be preserved long term in a refrigerator (4 °C). And it is possible to realize easy and safe transportation of sperm at an ambient temperature that requires neither liquid nitrogen nor dry ice. Furthermore, it has been demonstrated that the fertilizing ability of sperm cryopreserved or freeze-dried by the methods described in this chapter is well maintained during long-term preservation. This chapter introduces the latest protocols for cryopreservation and freeze-drying of mouse sperm, and the anticipated results of the fertilizing ability of these sperm preserved long-term.