Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
권호 표지

Chromosomal Analysis of Hanwoo Embryos by In Vitro Culture Condition

한우 체외 수정란의 체외 배양 조건에 따른 염색체 분석

  • Choi, S.H. (Animal Genetic Resources Station, NIAS, RDA) ;
  • Cho, S.R. (Animal Genetic Resources Station, NIAS, RDA) ;
  • Han, M.H. (Animal Genetic Resources Station, NIAS, RDA) ;
  • Kim, H.J. (Animal Genetic Resources Station, NIAS, RDA) ;
  • Choe, C.Y. (Animal Genetic Resources Station, NIAS, RDA) ;
  • Son, D.S. (Animal Genetic Resources Station, NIAS, RDA) ;
  • Chung, Y.G. (ET Biotech) ;
  • Kim, S.K. (Chungnam National University) ;
  • Sohn, S.H. (Jinju Industrial University)
  • 최선호 (축산과학원 가축유전자원시험장) ;
  • 조상래 (축산과학원 가축유전자원시험장) ;
  • 한만희 (축산과학원 가축유전자원시험장) ;
  • 김현종 (축산과학원 가축유전자원시험장) ;
  • 최창용 (축산과학원 가축유전자원시험장) ;
  • 손동수 (축산과학원 가축유전자원시험장) ;
  • 정연길 (ET 바이오텍) ;
  • 김상근 (충남대학교) ;
  • 손시환 (진주산업대학교)
  • Published : 2007.06.30
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Abstract

Antioxidants were well known to be essential supplements in the complex media and serve as a reservoir of oxygen. In this study, Hanwoo COCs (cumulus oocytes complexes) were matured and developed in L-cysteine-TCM199 and analyzed metaphase chromosome. Maturation rate of Hanwoo COCs were 73.4%, 94.6% in 0.1% PVA, 0.1 mM L-cysteine, respectively and showed significantly different between the treatments (p<0.05). Blastocyst formation were revealed 20.3%, 10.0% in 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA, respectively. There were no significant difference among treatment groups. Metaphase chromosome were showed 18.3%, 12.0% in 5% FBS-TCM199, 0.1 mM L-cysteine, respectively and analyzable chromosome were 6.1%, 4.0% and had no differences between the treated groups. In the case of in vitro developmental stages, metaphase chromosome were showed 18.3%, 12.0% in $4{\sim}16$ cells stage, 43.1%, 13.0% in morulae stage and 94.8%, 100.0% in blastocyst stage. These results suggested L-cysteine has beneficial role for in virto maturation and development in Hanwoo COCs.

항산화제는 산소의 저장고로서 무혈청 배양액에서 주요한 작용을 하며, 복합배지에서 유용한 첨가제로 알려져 있다. 따라서 본 연구는 한우 체외 수정란의 배양에 있어서 항산화제인 L-cysteine의 작용과 수정란의 발달 단계별 염색체의 분석을 통하여 체외 수정란의 배양 체계를 수립하고자 실시하였다. 한우 난포란의 체외 성숙은 0.1% PVA, 0.1 mM L-cysteine 첨가 시 체외 성숙율은 73.4%, 94.6%으로 각각 나타냈으며, 처리간에 유의적인 차이를 보였다(p<0.05). 체외 발달율은 20.3%, 10.0%로 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA 첨가구에서 각각 나타났으며, 처리간에 유의적인 차이는 보이지 않았다. 배양액의 종류에 따른 염색체 분석 결과는 중기상의 수정란은 18.3%, 12.0%를 보였으며, 분석 가능 수정란 수는 6.1%, 4.0%로 5% FBS+TCM199, 0.1mM L-cysteine 첨가구에서 각각 나타났고, 60, XX 2개, 60XY 1개가 5% FBS+TCM199 처리구에서, 60, XX 2개가 0.1 mM L-cysteine 처리구에서 확인되었고, 처리간에 유의적인 차이가 없었다. 수정란의 발달 단계별 염색체 분석 결과는 $4{\sim}16$세포기는 5% FBS-TCM199 배양액과 0.1 mM L-cysteine을 첨가한 배양액에서 18.3, 12.0%의 염색체 중기상을 확인할 수 있었고, 상실기에서는 43.1, 13.0%의 염색체 중기상을 보였으며, 배반포기의 경우는 94.8, 100.0%의 염색체 중기상을 보여 발달 단계가 진행될수록 염색체 중기 상이 많이 나타났다. 이상의 결과로 항산화제인 L-cysteine은 한우 난포란의 체외 성숙 및 발달에 중요한 인자임을 확인하였다.

Keywords

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