This study was carried out to evaluate the effect of culture methods on development of embryos with each developmental stage after heat shock in bovine oocytes. The results obtained were as fellows. 1. The culture method after heat shock on development of embryos was better drop-culture than co-culture. 2. The medium without amino acids were not effect of heat sock on development of embryos but it was in need of amino acid during formation of blastocyst.

the present study was carried out to develop a completely defined culture system and determine if high NaCl concentrations in defined (PVA added) or semi-defined (BSA added) medium is toxic to bovine embryos. Oocytes from slaughterhouse ovaries were matured and fertilized in vitro. After 30 h of insemination, only 2-cell stage embryos were selected and cultured for this experiment. The culture media used were as follows : TLP(114 mM of NaCl) + BSA (3 mg/ml), TLP + PVA (1 mg/ml), mTLP(96 mM of NaCl) + BSA, mTLP + PVA. Six to ten embryos were placed into a 301 drop of each medium and the embryos were examined at 10 day post-insemination without medium renewal. The experiment was replicated 4 times. All data were analyzed by chi-square. There were no significant differences among TLP-BSA, mTLP-BSA and mTLP-PVA in blastocyst development (21.6, 17.2 and 20.2%), respectively. Also, no differences were obtained in hatching rates (11.7, 9.9 and 12.2%), respecitively. However, there were significant differences between TLP-PVA (1.7% and 0.6%) and other group in blastocyst formation and hatching rates, respectively (p<0.01). Development of in vitro produced embryos cultured in BSA containing medium was not affected by high NaCl concentration, but in the completely defined medium, embryonic development was highly affected by NaCl. This study shows that reduced NaCl concentration in completely defined medium is beneficial for development of bovine pre-implantation embryos in vitro.

본 연구에서는 유기물 재료로 구득이 쉽고 수량이 풍부하며 재료의 균일성이 좋은 왕겨를 팽연화 하여 양액재배용 배지 재료 사용하기 위해 적합한 조건을 탐색해 보았다. 팽연화 왕겨의 물리성은 CEC 37.0cmol.kg-1, 가비중 0.19g.㎤, -0.1 bar 상태에서 보수력 271.0으로 펄라이트보다 우수하였으나 흡수속도는 펄라이트에 크게 떨어지는 경향이었다. 팽연화 수준이 높아짐에 따라 보수력, 흡수속도 등 수분관련 물성이 좋아지는데 반해 입도가 작아지고 부숙속도가 빨라지는 변화가 있었고 재배중 부숙이 진행됨에 따라 급액 15일 이후부터 pH가 상승되며 15~20일 사이에 NO3 부족이 심화되어 작물에 스테레스를 주는 것으로 판단되었다. ERH a처리에서 당도가 펄라이트에 비해 약 1.0 。Brix정도 높고 수량의 유의성은 없었으므로 틈새간극 8mm, 날높이 3mm에서 생산된 팽연화 왕겨의 물성이 양액재배용 배지 재료로 사용하기에 적정하였다.

적정 배양액의 선정, ITS의 첨가와 BSA의 농도조절 및 NaCl 농도의 조절을 통해 소 수정란의 무혈청, 체세포배제 배양체계를 확립하기 위하여 수행한 실험에서 다음과 같은 결론을 얻었다. 1. 배양액으로 CRlaa, TALP 및 SOF를 사용하여 발육률을 검토한 결과, 발육률의 유의적인 차이는 나타나지 않았다. 2. 배양액내의 고분자물질원으로 BSA, FBS 및 PVA를 첨가하여 사용한 결과 BSA 및 FBS 첨가군이 PVA 첨가군보다 유의적으로 높

The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.

본 실험은 단고추 자루식 장기 양액배지에 있어서 암면 대체배지를 개발코자 암면구를 대조구로하여 훈탄, 펄라이트, 훈탄(3) : 펄라이트(7), 훈탄(1) : 펄라이트(1), 출탄(7) : 펄라이트(3)를 두고, 이들 배지종류가 생육 및 수량에 미치는 영향을 시험한 결과를 요약하면 다음과 같다. 1. 초장과 엽수는 처리간 유의성이 인정되지 않았으나 훈탄(1) : 펄라이트(1)구에서 경경이 굵고 지상부 건물중과 근중이 무거웠으며 T/R율은 작았다. 2. 과장과 과수 역시 처리간에 유의성이 인정되지 않았으나 훈탄(1) : 펄라이트(1)와 훈탄(3) : 펄라이트(7)구에서 과장과 과수, 평균과중이 컸고, 수량도 많았다. 3. 식물체분석 결과 잎과 뿌리에 함유된 전질소와 인산은 훈탄(1) : 펄라이트(1)에서 많은 편이었다. 4. 수량은 수확 개시 4월 이후부터 계속 증가하여 7개월 후인 5월에 가장 많았고, 훈탄(1) : 펄라이트(1)구에서 수확 초기부터 후기까지 다른 처리구에 비하여 많았다.

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.