논문 상세보기

돼지 공여세포의 조건이 핵이식 수정란의 체외발달에 미치는 영향 KCI 등재

Effects of Donor Somatic Cell Conditions on In Vitro Development of Nuclear Transplanted Porcine Embryos

  • 언어KOR
  • URLhttps://db.koreascholar.com/Article/Detail/47323
구독 기관 인증 시 무료 이용이 가능합니다. 4,000원
한국동물생명공학회지 (구 한국수정란이식학회지) (Journal of Animal Reproduciton and Biotechnology)
한국동물생명공학회(구 한국수정란이식학회) (Journal of Animal Reproduction & Biotechnology)
초록

본 연구는 복제 돼지의 생산성 향상과 형질전환에 의한 대체상기용 복제 돼지 생산에 기여하기위한 기초연구로 공여세포의 조건, 핵이식 수정란의 융합 및 활성화와 체외발달에 미치는 각종 요인들은 조사하였다. 공여세포는 생후 10개월 된 Landrace 종으로부터 귀 세포조직(55mm)을 채취하여 0.05%의 trypsin과 EDTA가 첨가된 D-PBS로 세포를 분리하여 10% FBS가 첨가된 TCM-199 배양액으로 계대배양을 실시하여 사용하였다. 핵이식은

This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30 sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39, 5% in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r m site of donor cells (65.3%) than that of 25~30 m(42.5%) or 30 m(45.5%) cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.

저자
  • 홍승표(진주산업대학교 동물생명과학과)
  • 박준규(진주산업대학교 동물생명과학과)
  • 이명열(진주산업대학교 동물생명과학과)
  • 이지삼(진주산업대학교 동물생명과학과)
  • 정장용(진주산업대학교 동물생명과학과)