Recently the importance of cryopreservation to preserve the genes of extremely rare or endangered species has been increasing. Many spermatozoa can be obtained from one male. And strains can be established by in vitro fertilization and embryo transfer after thawing. Cryopreservation of spermatozoa is conducted in domestic animals and laboratory animals, but there has been almost no report about Mongolian gerbil spermatozoa. In this study, we examined the following: 1) Experiment 1 examined sperm viability and motility obtained with various concentrations of cryopreservation solution. 2) Experiment 2 compared the sperm viability obtained by two different methods of sperm collection. 3) Experiment 3 compared the effect of two different methods of sperm collection on sperm mobility and hyperactive movement. In Experiment 1, both the viability and motility obtained with mixtures of 1) 15% (w/v) raffinose and 5% (w/v) skim milk, 2) 10% (w/v) raffinose and 4% (v/v) glycerol, 3) 10% (w/v) raffinose and 4% (v/v) ethylene glycol were higher than those obtained with other mixture solutions. In Experiments 2 and 3, a mixture of 15% (w/v) raffinose and 5% (w/v) skim milk by the swim-up method was shown to result in high viability, motility and hyperactive movement. Moreover, hyperactive movement was highest after 2 hours in the control, whereas freezing spermatozoa showed their highest hyperactive movement after 1 hour.