In order to optimize the protocols for Cynomolgus Monkey(Macaca fascicularis)sperm cryopreservation,the effects of glycerol concentration [3%,5%,7% and 11%(v/v)] and equilibration time(10,30,60 and 90 min)on cryopreservation of spermatozoa of Cynomolgus Monkey were studied.A modified TTE(mTTE)dilution solution was used and the sperm motility,morphology,membrane integrity and acrosome integrity were examined before freezing and post-thawing.When 30 min equilibration was applied,the post-thawing motility and recovery rates was significantly higher than those when 90 min or 120 min equilibration(P<0.05).The effect of 30 min equilibration was also better than that of 60 min equilibration.The post-thawing motility and recovery rate of spermatozoa was significantly higher(P<0.05)for 3% and 5% glycerol groups than those for 11% glycerol group,and the effect of 3% and 5% glycerol was also better than 7% glycerol.The different glycerol concentrations and equilibration time showed similar effects on the membrane integrity,acrosomal integrity and malformation rate(P<0.05).The results suggest that addition of 3% or 5% glycerol and 30 min equilibration are beneficial for freezing Cynomolgus Monkey spermatozoa.