Effect Of The Main Factors On Sheep And Goat Embryo Production In Vitro

Many influence factors within IVF were experimented systematically in this study for the sake of establishing an acceptable IVF system on sheep and goat. The main contents of this study are: effect of different factors on sheep IVF technique and the effect of different factors on goat IVF technique.In the study of sheep IVF technique, the effects of SOFaa and CR1aa systems, the different ovary keep temperature (25℃, 37℃ ) and different methods of collecting oocytes, the different IVM time(21hs, 24hs, 27hs), the different supplemental dose of FSH, the different hormone combination strategy, the FBS heat-inactived or not and the supplement of 10% fresh PFF in IVM medium, the different IVF media and different sperm preparation methods, the influence of changing IVC medium during IVC with a SOFaa mixed gas system to the effect of sheep IVM/IVF/IVC were studied. The results shows, the cleavage rate and morula-blastocyst (M-B) development rate of SOFaa system are higher than those of CR1aa system(17.22%(47/273) and 19.15%(9/47) vs 10.68%(22/206) and 9.09%(2/22)). So the SOFaa system is better than CRlaa system on sheep IVF research in our Lab; as the transport time between 6-9hs, 25 ℃ is a suitable temperature to keep the ovary compared to 37 ℃; the aspiration method can obtain 1.43±0.36 good quality oocytes per ovary, a comparative number of good oocytes can also obtained by slicing the ovary aspirated(1.16+0.3), but even the two methods are synthetically used, the number of good oocytes is still sifnificantly lower than that of the direct-slicing method(2.59±0.82 vs 5.47±1.43, P<0.01), the cleavage rate of the direct-slicing method is significantly higher than that of aspiration method and the indirect-slicing method(59.93%(193/339) vs 43.90%(162/369) vs 38.86%(129/332), P<0.01), So the direct-slicing method is the best way to collecting oocytes; with SOFaa system, the better IVM time is 21hs-24hs; the supplement of FSH is essential to the IVM of sheep, the supplement of FSH in IVM medium can significantly increase the cleavage rate(P<0.01), the best dose of FSH is 20μg/ml for the sake of M-B development rate; LH and E2 has no obvious function to improve the cleavage rate and the M-B rate (P>0.05); the FBS heat-inactived have no obvious influence on the cleavage rate(P>0.05); The supplement of 10% fresh PFF to the...