Study On In Vitro Maturation And Parthenogenetic Activation Of Sheep Oocytes

Used the ovaries from slaughter house, sheep oocytes IVM(in vitro maturation), parthenogenetic activation, and embryos development from parthenogenetic activation were studied systematically. In different season, IVM oocytes of sheep with different concentration of FSH and LH were compared. Different temperature of sheep transported and different IVM time of sheep oocytes were compared. Effect of human HGF with different concentration on IVM of sheep oocytes was also compared. The result showed that different concentration of FSH and LH were needed for sheep oocytes IVM in different season Xinjiang. FSH 12.9ug/ml,LH0.86IU/ml is proper to sheep oocytes IVM in summer-autumn anestrous period. FSH 8.6ug/ml,LH0.57IU/ml is proper to sheep oocytes IVM in autumn oestrous period. and FSH 8.6ug/ml,LH0.86IU/ml is proper to sheep oocytes IVM in spring anestrous period. The appropriate time of sheep oocytes IVM was 24～26h. The appropriate sheep ovaries transport tempera- ture was 20～35℃. Human HGF restrain the sheep oocytes IVM. We designed a series of experiments using various activation methods with EH and electricity to determine the optimum activation efficiency of sheep oocytes. The result show that the optimum activation parameter of EH is 7%, 7min. The result of combined activation of EH and CHX is significantly better than each of them single activation. the optimum parameter of electric activation is 1.2kv/cm,3 pulse . Eliectric activation + A23187 + CHX was the best protocol to sheep oocytes activation (cleavage rate 69.8% and blastocyst rate 30%).Effects of different protein additive on sheep oocytes first cleavage after parthenogenetic activation were compared. Effects of NEAA with different concentration and different co-culture system on embryos development after parthenogenetic activation were compared too. The result showed that the first cleavage of sheep oocytes after activation was promoted by BSA + FBS. Morula/blastocyst development rate of sheep embryos from activation was improved significantly by adding 3.0% NEAA. alike-sequence culture system (TCM199+KL)+(mSOF+SLG)co-culture system was optimized to sheep embryos from activation culture in vitro.