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태아섬유아세포와 중간엽줄기세포를 이용한 체세포 복제돼지 생산 효율 KCI 등재

Cloning Efficiency of Porcine Somatic Cell Nuclear Transfer Compared with Fetal Fibroblasts and Mesenchymal Stem Cells

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동물자원연구 (Annals of Animal Resources Sciences)
강원대학교 동물자원공동연구소 (Institute of Animal Resources Kangwon National University)
초록

Somatic cell nuclear transfer (SCNT) in pigs has been used as a very important tool to produce transgenic for the pharmaceutical protein, xenotransplantation, and disease model and basic research of cloned animals. However, the production efficiency of SCNT embryos is very low in pigs and miniature pigs. The type of donor cell is an important factor influencing the production efficiency of these cloned pigs. Here, we investigated the developmental efficiency of SCNT embryos to blastocysts and full term development using fetal fibroblasts (FF) and mesenchymal stem cells (MSCs) to identify a suitable cell type as donor cell. We isolated each MSCs and FF from the femoral region and fetus. Cultured donor cell was injected into matured embryos for cloning. After that, we transferred cloned embryos into surrogate mothers. In term of in vitro development, the SCNT embryos that used MSCs had significantly higher in cleavage rates than those of FF (81.5% vs. 72%) (p<0.05), but the blastocyst formation rates and apoptotic cell ratio was similar (15.1%, 6.18% vs. 20.8%, 9.32%). After embryo transferred to surrogates, nine and nineteen clone piglets were obtained from the MSCs and FF group, respectively, without significant differences in pregnancy and birth rate (50%, 40% vs. 52.3%, 45.4%) (p>0.05). Moreover, there was no significant difference in the corpus hemorrhagicum numbers of ovary, according to pregnancy, abortion, and delivery of surrogate mothers between MSCs and FF groups. Therefore, the MSCs and FF are useful donor cells for production of clone piglets through SCNT, and can be used as important basic data for improving the efficiency of production of transgenic clone pigs in the future.

목차
ABSTRACT
Ⅰ. 서론
Ⅱ. 재료 및 방법
    1. 시약
    2. 난소의 채취 및 난자의 채란
    3. 난자의 체외성숙 및 체외배양
    4. 공여세포 준비
    5. 성숙난자의 선별과 핵이식
    6. 핵이식수정란의 세포사멸율 조사
    7. 체세포 복제돼지 생산
    8. 통계적 분석
Ⅲ. 결과
    1. 태아섬유아세포와 중간엽줄기세포를 공여세포로 한핵이식수정란의 비교
    2. 태아섬유아세포와 중간엽줄기세포를 공여세포로 한복제 돼지 생산 비교
Ⅳ. 고찰
Ⅴ. 요약
Ⅵ. 참고문헌
저자
  • 김효정(부산대학교 생명자원과학대학 동물생명자원과학과 대학원생) | Hyo-Jung Kim (Graduate Students, Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University)
  • 김민주(부산대학교 생명자원과학대학 동물생명자원과학과 대학원생, 한국생명공학연구원 연구원) | Min Ju Kim (Graduate Students,Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University, 4Researcher, Futuristic Animal Resource & Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang 28116, Korea)
  • 박수현(부산대학교 생명자원과학대학 동물생명자원과학과 대학원생) | Soo-Hyun Park (Graduate Students,Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University)
  • 주예은(부산대학교 생명자원과학대학 동물생명자원과학과 대학원생, 한국생명공학연구원 연구원) | Ye Eun Joo (Graduate Students,Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University, 4Researcher, Futuristic Animal Resource & Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang 28116, Korea)
  • 허창기((주)크로넥스 연구원) | Chang-Gi Hur (Researcher, CRONEX, Inc., Hwaseong 18525, Korea)
  • 심보웅(한국생명공학연구원 연구원) | Bo-Woong Sim (Researcher, Futuristic Animal Resource & Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang 28116, Korea)
  • 김명후(부산대학교 생명자원과학대학 동물생명자원과학과 교수)
  • 서자겸(부산대학교 생명자원과학대학 동물생명자원과학과 교수) | Jakyeom Seo (Professor, Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University)
  • 김병우(부산대학교 생명자원과학대학 동물생명자원과학과 교수) | Byeong-Woo Kim (Professor, Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University)
  • 조병욱(부산대학교 생명자원과학대학 동물생명자원과학과 교수) | Byung-Wook Cho (Professor, Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University)
  • 신택순(부산대학교 생명자원과학대학 동물생명자원과학과 교수) | Taek- Soon Shin (Professor, Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University)
  • 조성근(부산대학교 생명자원과학대학 동물생명자원과학과 교수) | Seong-Keun Cho (Professor, Department of Animal Science, College of Natural Resources & Life Science, Life and Industry Convergence Research Institute (LICRI), Pusan National University) Corresponding author