The Research On Vitro Cultivation And Parthenogenetic Activation Of Oocytes In Sow

The focuses of this experiment are to set up and optimize the sow oocytes in vitro maturation and cryopreservation,parthenogenetic activation,which lay the foundation for further study of the laboratory.The research for sow in vitro maturation and cultivation helps to reveal the matural mechanism on the sow oocytes,which provides a evidence for the foundation on sow embrydeterminedlyo in vitro production technology system.The results show that:1.It has set up a maturation on sow oocytes system and determined the working concentration for some hormones from the matural liquid in sow oocytes,finally the prescription for matural liquid is that TCM-199(Earle's)+0.1%PVA+3.05 mM D-glucose +0.91 mM sodium pyruvate +75 mg/mL penicillin + 50 mg/mL streptomycin + 0.57 mM cysteine + 20IU/ml PMSG + 20IU/ml hCG + 30 ng/ml EGF + 10%pFF;2.By comparation the effect on sow oocyte in vitro maturation for adding different concentrations of EGF,results show that it can improve matural rate of the sow oocyes significantly by adding EGF,and the matural rate of oocyes will be improved continually as increasing EGF.the capacity of EGF is 50 ng/ml,the matural rate of oocytes will be 78.1%;3.We found that it would improve the matural rate of the sow oocyes with adding the concentration of PFF appropriately by study,and the definitively working concentration was 10%in this experiment;4.By studying the influence of time for transporting ovary and the temperature of preservation on matural rate of oocytes,results show that,4h or less,temperature controlled at about 35℃will be better on sow oocytes in vitro maturation,and the matural rate of oocytes will be more than 75%;5.The experimental comparison,we found that the matural rate on oocytes from the ovary in follicular phase and in luteal phase was no significant difference;6.By comparison the effect on sow oocytes in vitro maturation in different maturation time,results showed that the matural rate would be increased as time was increased,and matural rate could be 78.6%in 48 hours;However,at 48h after parthenogenetic activation on the cleavage rate has decreased; 7.Through experimental studies,the parthenogenetic activation system on sow oocyes is lonomycin +6- DAMP + CB,the best time for oocytes at the role of 6-DAMP is 4h;8.The experimental comparison,Vitrification for cryopreserving sow oocytes was more effective,and cryoprotectants selected EG or EG + DMSO mixed group;9.The experimental comparison,the Frozen effectiveness on oocytes in GV period was significantly higher than those in MⅡperiod,and there were significant differences between them;10.The experimental comparison,the steps for frozing was the 6-step frozen,and thaw steps was 4-step thawing.