The Research On Mature In Vitro And Pathenogenetic Activation Of Porcine Oocytes

The objective of this paper was trying to establish an optimum mature system to improve mature efficiency,and find a proper method of pathenogentic activation of porcine oocytes matured in vitro.In vituo mature (IVM) rate and Pathenogentic activation rate of porcine oocytes,at present,are still low,which become a barrier to progress in research of porcine embryonic engineering. The effect of different oocytes , different culture medium of COCs, different culture medium in mature system and pathenogentic activation methods on porcine oocyte development were investigated in this study .The results were as follows:The IVM rate of porcine oocytes derived from follicles in diameters (<3mm,3-6mm,> 6mm) were increased gradually(65.5﹪,81.6﹪,81.8﹪,p<0.05)The proportion of class A cumulus-oocytes was higher than the proportion of class B cumulus-oocytes in the expanded cumulus rat(e100%,52.8%,P<0.05),,but there were no significant differences in the IVM rates(81.6%,79.3% P>0.05).Both of the class A and B can get the higher rate of the IVM than the class C.Porcine oocytes were cultured in NCSU-23,TCM199, There were no significant differences in the IVM rate(80.6% , 79.8% ,p>0.05). Porcine cumulus- oocyte complexes(Cocs) were cultured in medium NCSU -23 for 36 h, 40 h, 44 h, 48 h. Rate of PBI were 7.3%,60.9%,83%, 63.4%, there were the significantly different(P<0.05). Therefore, the best time for adapted to culture porcine oocytes in vitro is 44 h , and the rate of maturation is to 83%.The porcine oocytes were activated by different electric field strength (100V/mm, 150V/mm, 200V/mm, 250V/mm), different time(20μs, 30μs, 40μs, 60μs, 80μs),different electric pulsing number (1time,2times,3times). The porcine oocytes were also activated by electrical pulses(200V/mm, 60μs , 1time) combined with chemical activation(CHX, 6-DMAP and CHX+6-DMAP). 200 V/mm of electric field strength and 60μs of electric field duration one electric pulsing can get the higher rate of oocyte activation,cleavage and parthenogenetic embryo. There were no significant differences in the cleavage rates(69.17%,82.31% and 79.67%) and blastocyst rates(24.17%,39.46% and 39.84%) after parthenogenetic activation using CHX,6-DMAP and CHX+6-DMAP.Porcine parthenogenetic embryos were cultured in NCSU-23,TCM-199和B2. There were no significant differences in the cleavage rates(82.05%,81.5%,82.35%), porcine parthenogenetic embryos were cultured in B2 can get the higher rate of blastocyst rate(40.48%)than in TCM-199(P<0.05)...