PORCINE EMBRYO PRODUCTION BY IN VITRO FERTILIZATION OF OOCYTES MATURED IN VITRO USING NCSU-37 SUPPLEMENTED WITH 10 PORCINE FOLLICULAR FLUID (PFF)

摘要

Porcine embryo in vitro production (IVP) systems which generate viable embryos as well as in vivo embryos will contribute to research in reproductive physiology, agriculture and biotechnologies. These systems included in vitro maturation (IVM) of oocytes and in vitro fertilization (IVF) and their subsequent in vitro culture (IVC), have been carried out by the researchers in National Institute of Animal Husbanry (NIAH). The study had been applied IVP procedures of Dr. Kazuhiro Kikuchi. We collected cumulus-oocyte complexes (COCs) of porcine ovaries from the slaughterhouses. Porcine COCs were matured in IVM1 at 38.5℃ in humidified condition of 5% CO2 in air for 20-22h, and then cultured in IVM2 for 24h. Only the oocytes with expanded cumulus cells were used for in vitro fertilization. The boar frozen semen was used for Porcine IVF procedure. The co-incubation for fertilization was carried out for 3 hours at 38.5℃ under 5% CO2. They were cultured in IVC1 medium supplemented with pyruvate and lactate from day 0 to day 2 then in IVC2 medium supplemented with glucose from day 2 to 6 under 5% CO2 at 38.5℃. There were some our results. In total 1392 COCs were matured in IVM1 medium and IVM2 after 40-44h. Collections of 915 (65.73%) oocytes with expanded cumulus cell were used for in vitro fertilization. The fertilization rates were 63.49%,The blastocyst formation rates on day 6 were 13.59%. These results show that we can produce porcine embryos in Vietnam.