The study was conducted to determine the optimal hormone and glucose levels during the in vitro culture of bovine oocytes matured and fertilized in vitro for blastocyst development. Oocytes matured in TCM 199 + 10% FCS + hormones and glucose were fertilized in vitro in a TALP medium with swim up separated and heparin-treated epididymal cauda spermatozoa. Oocytes were cultured for 2~5 days in synthetic oviduct fluid medium (SOFM) supplemented with 10% FGS and with different hormone and glucose levels, and further cultured 5 days same medium in SOFM. The results are summarized as follows : The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + estradiol, PMSG + estradiol 0 to20 hours after insemination were 88.0% and 81.8%, 82.6% and 68.4%, 80.0% and 75.0%, 80.0% and 65.0%, 77.3% and 64.7%, respectively. The in vitro maturation and penetration rates of porcine oocytes cultured in TCM 199 media containing PMSG, hCG, PMSG + hCG, hCG + estradiol, PMSG + estradiol 20 to 40 hours after insemination were 92.0% and 87.0%, 92.0% and 82.6%, 91.3% and 81.0%, 85.2% and 73.9%, 87.5% and 81.0%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose lelvels 0~3 days after insemination were 31.5~48.1% and 10.0~16.7%, respectively. The cleavage and in vitro developmental rates to blastocyst of porcine oocytes cultured in TCM 199 media containing 0.05 mM, 0.10 mM, 0.30 mM, 0.50 mM, 1.00 mM, and 3.00 mM glucose levels 4~8 days after insemination were 30.0~53.8% and 8.7~19.2%, respectively. The cleavage and in vitro developmental rates to blastocyst were higher in TCM 199 media containing various glucose levels 0~3 days after insemination than 4~8 days.

This research was conducted to investigate the interrelationship among methods of injection of PMSG-hCG to the number of ovulated eggs, percentage of matured oocytes and in vitro fertilization using out-bred ICR mice. The results obtained are as follows, 1) The optimurn dose was 5 IU for both PMSG and hCG, while the number of ovulated eggs was 428, percentage of M II was 73% and in vitro fertilization rate was 81 %. 2) The optimum injection interval of PMSG-hCG was 48 hours, while the number of ovulated eggs was 48 8, percentage of M II was 80% and in vitro fertilization rate was 81%. 3) The optimum time for collecting eggs was between 16 and 18 hours after hCG injection, while the numbers of ovulated eggs were 448, 427 and 437 in 14,16 and 18 hours after hCG injection respectively, and percentages of M II were 79 and 81 %, and in vitro fertilization rates were 81 and 80% in 16 and 18 hours after hCG injection, respectively. 4) The repeat of superovulation decreased with the number of ovulated eggs, percentage of M II and in vitro fertilization rate, than in control. But it was recovered by increasing the repeat interval.