생쥐 복제수정란 발달에 있어서 난자공여 생쥐 연령과 미세조작 배양액의 영향

The Effect of Oocyte Donor Age and Micromainpulation Medium on the Development of Mouse Cloned Embryos

  • 김동훈 (국립축산과학원 동물바이오공학과) ;
  • 이윤수 (동경대학교 의과학연구소) ;
  • 오건봉 (국립축산과학원 동물바이오공학과) ;
  • 황성수 (국립축산과학원 동물바이오공학과) ;
  • 임기순 (국립축산과학원 동물바이오공학과) ;
  • 박진기 (국립축산과학원 동물바이오공학과)
  • Kim, Dong-Hoon (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Youn-Su (Laboratory of Stem Cell Therapy, Center for Experimental Medicine, Institute of Medical Science, University of Tokyo) ;
  • Oh, Keon-Bong (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Hwang, Seong-Soo (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Im, Gi-Sun (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Park, Jin-Ki (Animal Biotechnology Division, National Institute of Animal Science, RDA)
  • 투고 : 2011.09.02
  • 심사 : 2011.09.09
  • 발행 : 2011.09.30

초록

This study was conducted to examine the effect of oocyte donor age and micromanipulation medium on the development of mouse cloned embryos receiving cumulus cells. Mouse oocytes were obtained from 6 to 11 week-old mice BDF1 female mice(experiment 1) and cumulus cells were used as donor cells. Micromanipulation procedures for nuclear transfer(NT) were performed in FHM, M2 or Hepes-buffered TCM199(TCM199) medium(experiment 2). After nuclear transfer, the reconstructed oocytes were activated by 10 mM $SrCl_2$ in Ca-free CZB medium in the presence of 5 II ${\mu}$g/ml cytochalasin B for 5 h and cultured in KSOM medium for 4 days. In experiment 1, the survival rate of oocytes after injection of cumulus cells were significantly(p<0.05) lower in oocytes from 6~7 week-old mice(53.3%) than in oocytes from 8~9(80.9%) and 10~11 week-old mice(77.1%). In experiment 2, the survival rate of oocytes after cell injection were significantly(p<0.05) higher in FHM and M2 medium(71.7% and 76.9%) than in TCM199 medium(51.2%). The activation rates of cloned embryos were not different among the micromanipulation media. However, the embryos developed to blastocyst stage were significantly(p<0.05) higher in FHM medium(13.9%) than in M2 and TCM199 medium(0.0% and 0.0%). In conclusion, the present study suggest that oocytes from above 8 week-old mice are superior to oocytes from 6~7 week-old mice as a source of recipient cytoplasm and FHM is superior to M2 and TCM199 as a micromanipulation medium for mouse somatic cell cloning.

키워드

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