The Research Of Porcine Oocytes In Vitro Maturation And Production Of Embryos By Intracytoplasmic Sperm Injection

Successful offspring production after a new technique which was intracytoplasmic sperm injection(ICSI)combined with Sperm mediated Gene Transfer(SMGT)had been reported in many laboratory animals.With the development of the embryo biotechnology,the demands for oocyte quality derived from maturation in vitro are increasingCurrently obtained by in vitro maturation of oocytes cytoplasmic maturation is less than the body of mature oocytes,there are many influence factors affects the quality of the in vitro maturation,such as time control of in vitro maturation,in vitro maturation medium different additives,oocyte collection method and oocyte quality and so on.This experiment study how to pick out the right oocyte for in vitro maturation in most oocytes.Procine oocytes for materials,pig oocytes in vitro fertilization and other related issues were studied.Also explored intracytoplasmic sperm injection preparation embryonic.Experimental content:According to the number of layers and the interity of the cumulus cells,oocytes into A,B,C three groups.And were cultivated in vitro maturation,After in vitro maturation of oocytes 44h,we observed the first polar body.The test results:A,B,C three groups the first polar body rate were 68.94%,47.94%,31.65%.The results showed that the rate of the first polar body rate was significantly higer than B and C.A and B was extremely significantly different(P<0.01).A and C was extremely significantly different(P<0.01),B and C was extremely singnificantly different(P<0.01).A,B,C three groups of oocytes parthenogenetic activation experiments.After 24-48h observed cleavage situation and 5-6d observed blastocyst,the results of the experimental for the cleavage rate was 87.72%,82.38%,52.94%.A and B cleavage rate was not significantly different(P>0.05).A and C was extremely significantly different(P<0.01).B and C was extremely significantly different(P<0.01).After 5-6d we observed blastocyst rates was 10.96%,6.21%,2.88%.A group blastocyst rate was higer than B and C.A blastocyst rate and C blastocyst was singnificantly different(P<0.05),B blastocyst rate was higer than C.blastocyst rate was singnificantly different(P<0.05).Different levels of mature oocytes after in vitro fertilization.A group oocytes blastocyst rate(10.61%)was significantly higher than B(5.34%)and C(3.58%).A and B blastocyst rate was extremely significantly different(P<0.01).B and C blastocyst rate was not significantly different(P>0.05).This results suggest that cumulus cells have a great impact on the embryos.?We conducted ICSI preparation of pig embryos experiments,We successfully prepared six fertilized blastocysts,this test foundation for transgenic pig reserach.Conslusion:cumulus cells have a significant impact on the developlement of in vitro maturation and embryos fertilized.Three or more layers wrapped dense cumulus-oocyte complexes in vitro maturation is the best choose.we use ICSI technology successful prepared six fertilized blastocysts.