Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Effects of Some Factors on In Vitro Production of Embryos from Antral Follicle-Derived Porcine Oocytes I. Effects of Maturation Media on In Vitro Maturation, Fertilization and Development

돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향 I. 체외성숙, 체외수정, 체외발달에 대한 체외성숙 배양액의 영향

  • Yeon, S.-H. (Animal Genetic Resources Station, National Livestock Research Institute, R.D.A.) ;
  • Choi, S.-H. (Animal Genetic Resources Station, National Livestock Research Institute, R.D.A.) ;
  • Kim, C.-D. (Animal Genetic Resources Station, National Livestock Research Institute, R.D.A.) ;
  • Son, D.-S. (Animal Genetic Resources Station, National Livestock Research Institute, R.D.A.) ;
  • Han, M.-H. (Animal Genetic Resources Station, National Livestock Research Institute, R.D.A.) ;
  • Lee, K.-S. (Division of Animal Science and Resources, Chungnam National University)
  • 연성흠 (논촌진흥청 축산연구소 가축유전자원시험장) ;
  • 최선호 (논촌진흥청 축산연구소 가축유전자원시험장) ;
  • 김종대 (논촌진흥청 축산연구소 가축유전자원시험장) ;
  • 손동수 (논촌진흥청 축산연구소 가축유전자원시험장) ;
  • 한만희 (논촌진흥청 축산연구소 가축유전자원시험장) ;
  • 이규승 (충남대학교 동물자원학부)
  • Published : 2004.08.01
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Abstract

This study was carried out to examine the effects of maturation media on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-37 (mNCSU-37), modified NCSU-23 (mNCSU-23), or TCM-199 supplemented with 10% porcine follicular fluid (pFF). Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium(mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU-23. The results are as follows. 1. In the result of IVM, the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different among the media, though numeric value of them were slightly lower in TCM-199 than in mNCSU-37 or in mNCSU-23. 2. In the result of IVF, though the rate of sperm penetration was not significantly different among the maturation media, the percentage of oocytes with male pronucleus (MPN) of ones matured in mNCSU-37 (88.0%) was significantly higher than in TCM-199 (71.1%) (p<0.05). 3. In the result of IVD, the percentage of cleaved oocytes of ones matured in mNCSU-37 (52.3%) or in mNCSU-23 (53.7%) was significantly higher than in TCM-199 (43.1%) (p<0.05), but the rate of blastocysts at day 6 was not significantly different among the maturation media, though putative embryos from oocytes matured in mNCSU-37 or in mNCSU-23 were developed more than in TCM-199. These results suggested that mNCSU-37 or mNCSU-23 was more appropriate than TCM-199 as IVM medium for porcine immature oocytes.

본 연구는 보다 안정된 돼지 체외수정란 생산시스템 확립을 목적으로 체외성숙배양액 mNCSU-37, mNCSU-23 및 TCM-199 가 각각 미성숙 난포란의 체외성숙, 체외수정, 체외발달에 미치는 영향을 구명하기 위하여 수행하였다. 도축돼지의 난소에서 채취한 COCs를 10% pFF가 포함된 각각의 성숙배양액에서 최종동도가 1${\times}10^5/m{\ell}$ 의 농도로 체외수정 시킨 다음, NCSU-23 에서 체외발달을 유도한 결과는 다음과 같다. 1. TCM-199에서 성숙시킨 난자가 mNCSU-37 이나 mNCSU-23에서 성숙시킨 것보다 난핵포 붕괴율과 핵 성숙율에서 다소 낮은 경향을 보였으나 유의적인 차이는 아니었다. 2. 정자 침투율은 성숙배양액간 유의적인 차이를 나타내지 않았으나 웅성정핵 형성율은 mNCSU-37 에서 성숙시킨 난자가 88.0%로 TCM-199 에서 성숙시킨 것의 71.1%보다 유의적으로 높았다.(p<0.05). 3. 난분할율은 mNCSU-37(52.3%)이나 mNCSU-23(53.7%)에서 성숙시킨 난자가 TCM-199(43.1%)에서 성숙시킨 난자보다 유의적으로 높았다(p<0.05). 배반포발달율도 mNCSU-37이나 mNCSU-23에서 성숙시킨 것이 TCM-199에서 성숙시킨 것보다 다소 높았지만 유의적인 차이는 아니었다. 이상의 결과로 보아 돼지 미성숙 난포란 유래의 체외수정란 생산을 위한 체외성숙배양액으로 TCM-199보다 mNCSU-37이나 mNCSU-23이 적합한 것으로 사료된다.

Keywords

References

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