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질환모델마우스 생산을 위한 체세포핵이식방법 개선; I. 탈핵 및 재조합난자 생산기법 최적화
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  • 질환모델마우스 생산을 위한 체세포핵이식방법 개선; I. 탈핵 및 재조합난자 생산기법 최적화
  • Improvement of Somatic Cell Nuclear Transfer Technology for the Production of Disease Model Mouse: I. Optimization of Oocyte Enucleation and Reconstruction
저자명
전수현,심호섭,정형민,이병천,이은송,고정재,신태형,박찬,황우석
간행물명
韓國受精卵移植學會誌
권/호정보
2000년|15권 3호|pp.247-253 (7 pages)
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한국수정란이식학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

This study was undertaken to optimize enucleation and reconstitution methods for the production of cloned mice by somatic cell nuclear transfer Outbred ICR mouse oocytes at the metapahse- II stage were retrieved from female mice superovulated by PMSG and hCG. In Experiment 1, oocytes were enucleated in medium supplemented with cytochalasin B (CCB) of 3 levels (0, 7.5 or 15 $mu extrm{g}$/mL), and higher rate of encleation was obtained at 7.5 and 15 $mu extrm{g}$/mL than at $mu extrm{g}$/mL. In Experiment 2, oocytes enucleated in 7.5 $mu extrm{g}$/mL CCB-containing medium were reconstituted with different types of somatic cell by following methods; 1) cumulus cells by direct cell injection, 2) cumulus cells by electric fusion (1.25 kV/cm, 2 pulses for each 70 $mutextrm{s}$) or 3) STO cells by the electrofusion. Electrofusion of STO cells with enucleated oocytes yielded the greatest (P<0.05) rate of reconstitution without lysis (76%) than any other combinations. Although significant decrease in the rate of somatic cell introduction was found, the electrofusion of cumulus cells yielded better rate of reconstitution than direct injection (0 vs. 18%). In Experiment 3, the duration of electric stimulation for the fusion was changed to either 50 $mutextrm{s}$ or 90 $mutextrm{s}$, but no significant improvement of reconstitution efficacy was obtained. In conclusion, this study showed that ICR mouse oocytes could be used for the production of reconstituted oocytes and a fusion method of 1.25 KV/cm with 2 pulses using 570 cell was the optimal.