Effects Of Freezing And Cold Shock On The Biological Characteristics And Mitochondrial Membrane Potential Of Spermatozoa

Sperm cryopreservation is the key technology to save sperm,In vitro fertilization and animal breed conservation.It has become a key content in the pig assisted reproductive technology in recent years.It can conduct long-term semen cryopreservation,transportation and IVF.The research explored freezing and cold shock sperm the biological characteristics,mitochondrial membrane potential,sperm adhesive property,the effect of IVF rate.Using the main method of microbial dynamic (static) state image detection system, coomassie brilliant blue staining method,low hypo-osmotic swelling test(HOST),JC-1(Mitochondrial membrane potential assay kit with JC-1) staining method.the microscopic examination of pig sperm adhesive property, and so on. We have following results after doing a lot of experiment:1.Using CAS A test the quality parameters of-196℃ reezing and thawing sperm,it is significantly higher than that the-80℃ freezing and thawing sperm and the cold shock sperm;2.In the IVF test 2-cell developmental rate of-196℃ freezing and thawing sperm is better than-80℃ freezing and thawing sperm;3.Though the HOST test,show that the tail crimp ratio and acrosome integrity of-196℃ freezing and thawing sperm is higher than-80℃ freezing and thawing sperm;4.The adhesion of oocytes-196℃ freezing and thawing sperm is better than-80℃ freezing and thawing sperm and cold shock sperm;5. Sperm mitochondrial membrane potential (△ψm) in the group of 25℃,-80℃,-196℃ that add β-entrapped cyclodextrin complex (CLC) are all higher than the group without CLC, and the △ψVm value is highest in the group add CLC at the temperature-196℃.