| The cynomolgus macaque is one of the important non-human primate in biomedical research,which is very close to human in physiology,genetics and behavior.The preservation of germplasm resources of non-human primates is very important for the protection of genetic diversity.Sperm cryopreservation of plays an important role in the protection of genetic biodiversity of endangered species,Ex-situ conservation and artificial propagation.Because primate sperm particularly sensitive to low temperature environments,the optimal cryopreserving protocol especially based on egg yolk free freezing meium is still not established,optimization is needed to improve the cryosurvival of cynomolgus sperm after freezing and thawing.In this study,based on the egg-yolk-free sperm freezing medium with type III antifreeze protein,the sperm collected from cynomolgus macaques was used to explore the effect of freezing and warming rates by different carriers and different concentration and combination of antioxidant superoxide dismutase and N-acetyl-L-cysteine on the cryosurvival of cynomolgus macaque sperm.The results showed that the desired cryosurvival of sperm was achieved at a freezing rate at-21.3 °C/min and a rewarming rate at 198.9 °C/min cryopreservation with 1.0 m L cryovival.However,the post-thaw sperm motility,acrosomal integrity and mitochondrial membrane potential keep decreasing following the in vitro culture for0,1,2 and 4 h.The combination of 1 m M NAC and 1000 U/m L SOD showed the highest cryosurvial rate of cynomolgus macaque sperm after thawing.This study provides an experimental basis for further optimization and protection of cynomolgus monkey sperm cryopreservation methods,and will be benifical for the preservation of germplasm resources of other nonhuman primates. |
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