Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Development of Chimeric Embryos Aggregated with Blastomeres from Parthenogenetic and in vitro Fertilized Bovine Embryos

소의 단위 발생란과 체외수정란 유래의 할구 응집에 의한 키메라 수정란(Chimeric Embryo)의 발달

  • E. H. Yeao (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Kim, Y. S. (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • Lee, S. L. (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • T. Y. Kang (Department of Veterinary Medicine, College of Agriculture and Life Sciences, Cheju National University.) ;
  • D. O. Kwack (Division of Science Education, Gyeonsang National University.) ;
  • Lee, H. J. (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University) ;
  • S. Y. Choe (Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University)
  • Published : 2003.04.01
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Abstract

발생학에서 키메라(chimera)는 2개 이상의 다른 유전자형의 세포, 또는 다른 종의 세포로부터 만들어진 1개의 생물개체를 뜻하는 말로, 이는 초기 수정란의 발달과 포유류의 분화를 연구하는데 이용되고 있다. 키메라를 만드는 방법에는 할구와 내세포괴를 응집시키는 방법과 배반포 내에 여러 종류의 세포를 주입하는 방법이 있다 본 실험에서는 서로 다른 두 가지 방법의 활성화 처리법, 즉, ionomycin 처리 후 Cycloheximide (CHX) 또는 6-Dimetylaminopurine (6-DMAP)에 따른 단위 발생란의 분할과 단계적인 발달율을 살펴 보고자 하였으며, 서로 다른 방법에 의해 생산된 단위발생란 유래의 할구와 체외수정란 유래 할구를 응집하여 키메라 수정란(chimeric embryo)를 만든 후 체외수정란과 발달율을 비교해 보았다. 도축장 유래의 난소에서 난자를 채취하여 체외에서 22~24시간 성숙시킨 후 난구세포를 제거하고 metaphase II 단계의 난자를 5 $\mu$M ionomycin에 4분간 처리한 후, 10 $\mu$g/ml CHX/5 $\mu$g/ml cytochalasin B (CCB)에 5시간 또는 1.9 mM 6-DMAP에 4시간 처리하여 분할율과 배반포기 발달율을 비교 조사하였다. 난자 분할율에서는 체외수정란과 6-DMAP처리 단위 발생란에서 각각가 83.7 와 85.5%로 CHX/CCB 처리 단위발생란의 57.9%보다 유의적으로 높게(P<0.05) 나타났으며, 배반포기 발달율에 있어서는 체외수정란의 발달율이 27.8%로 6-DMAP처리 활성란 12.3%와 CHX/CCB 처리 활성란 5.3%보다 유의적으로 높게 (P<0.05) 나타났다. 키메라 수정란(chimeric embryo)은 서로 다른 두 가지 처리에 의해 생산된 단위발생란의 할구 2개와 체외수정란 유래의 할구 2개를 빈 투명대 내에서 응집시켜 제조하였다 빈 투명대 내에 키메라 수정란(chimeric embryos)의 8 세포기까지의 발달율은, 체외 수정란 할구 2개와 CHX/CCB 처리에 의한 할구 2개를 응집한 그룹은 46.1%, 체외 수정란 할구와 6-DMAP 유래 할구 2개를 응집한 그룹은 52.8% 였으며, handled control은 54.7%로 체외 수정란 77.7%에 비해 유의적으로 낮게(P<0.05) 나타났다. 배반포기까지의 발달율은 체외 수정란과 CHX/CCB에 의해 생산된 키메라 수정란(chimeric embryo)은 12.8%, 체외 수정란과 6-DMAP에 의한 키메라 수정란(chimeric embryo)은 18.8%로 handled control의 21.4%에 비해 유의적으로 낮았으며(P<0.05), 이들 키메라 수정란(chimeric embryos)은 체외 수정란의 34.9%에 비해 유의적으로 낮게(P<0.05)나타났다. 6-DMAP 처리 단위발생이 유기된 수정란 할구 2개와 체외수정란의 할구 2개의 응집에 의한 키메라 수정란(chimeric embryos)의 발달율이 CHX/CCB와 체외수정란의 응집에 의한 키메라 수정란(chimeric embryos)에 비해 다소 높게 나타났으나, 유의적인 차이는 없었다. 본 실험의 결과 서로 다른 방법에 의한 단위 발생란 유래의 할구와 체외 수정란 유래의 할구가 응집에 의한 재조합이 가능하였고 이들을 체외에서 배양하여 배반포기의 수정란까지 발달시켰다.

Keywords

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