Study On The IVF And OPS Vitrificational Cryopreservation Of Porcine Oocyte

In this investigation, we study the effect of season, FCS, pFF, insulin and estradiol on pig oocytes in vitro maturation(IVM); the effect of different cyroprotectant, methods of treatment for oocytes (treated with CB or centrifuge) and oocytes in different stage on OPS (open pulled straw) cryopreservation; the effect of different medium, concentration of caffeine on pig in vitro fertilization (IVF); the effect of protein,insulin and glutamine on development of pig preimplantation embryos in vitro.Result, 1) maturation rates of pig oocytes from IVM in Spring and Autumn was higher than other season (P<0.05). Pig oocytes from ovary in follicular phase had higher maturation rates than luteal phase. After adding 10%FCS,10%pFF,20%FCS and 10%FCS+10%pFF to maturation medium, the cleavage rates had no significant different (P>0.05). Maturation medium with insulin and estradiol can improve maturation and cleavage. 2) cryoprotectant with 5%PVP and ethylene glycol(EG) improved the freezing result, oocytes in GV phase had better freezing result than other phase, treated oocytes with CB improved cleavage after thawing, but centrifuging oocytes before cryopreservation achieved opposite results for embryo in vitro development. 3) Zygotes in mBO with 5mM caffeine improved fertilization, adding FCS to medium after 24h development, insulin and glutamine all improved pig embryos in vitro development.Conclusion in our study, 1) NCSU23 with 10% FCS as basic maturation medium, adding hormone in the first 24h to medium, there was a better maturation rates during 48h in vitro maturation. 2) We used cryoprotectant with 5%PVP +EG and five steps OPS cryopreservation. After in vitro maturation 12h, these oocytes (GV phase) take more advantage of cryopreservation and had the best development after thawing in our research. 3) mBO with 5mM caffeine as fertilization medium had the best fertilization result.