항산화제와 Growth Factors 첨가가 돼지 체외수정란의 체외발육에 미치는 효과

Effect of Antioxidants and Growth Factors on the Development of Porcine IVM/IVF Embryos

  • 최영진 (강원대학교 동물자원과학대학) ;
  • 박춘근 (강원대학교 동물자원과학대학) ;
  • 정희태 (강원대학교 동물자원과학대학) ;
  • 김정익 (강원대학교 동물자원과학대학) ;
  • 박동헌 (강원대학교 동물자원과학대학) ;
  • 장현용 (강원대학교 동물자원과학대학) ;
  • 양부근 (강원대학교 동물자원과학대학)
  • 발행 : 2002.06.01

초록

본 연구는 돼지의 미성숙 난포란을 체외에서 성숙, 수정시킨 후, 생산된 체외수정란을 NCSU23 체외배양액에 항산화제(NAC, ebselen 및 glutathione)와 성장인자(EGF와 PDGF)의 첨가가 돼지 체외수정란의 체외 발육에 미치는 영향을 검토하였다. NAC을 0, 0.5, 1.0 및 2.0mM 첨가하여 체외배양한 결과, 상실배기 이상 발육된 체외발육성적은 1.0mM과 2.0mM 첨가구가 여타구보다 높은 체외발육성적을 나타냈다. 한편, ebselen을 0, 5, 10 및 25$\mu$M을 첨가하여 배양한 경우 상실배 이상 발육된 체외발육 성적은 각각 17.9, 22.5, 36.9 및 10.8%로서 ebselen 10$\mu$M 첨가구가 여타구에 비해 다소 높은 체외발육성적을 얻었으나 통계적 유의차는 인정되지 않았다(P>0.05). 또한 glutathione을 0, 50, 100 및 200$\mu$M 첨가시 상실배기 이상 발육된 체외발육성적은 각각 21.8, 17.4, 28.3 및 15.8%로서 100$\mu$M 처리구가 여타구보다 다소 높은 체외발육성적을 얻었으나 통계적 유의차는 인정되지 않았다(P>0.05). 돼지 체외수정란의 체외배양시 체외배양액에 growth factor의 첨가배양이 돼지수정란의 체외발육에 미치는 영향을 검토한 결과, NCSU 23배양액에 EGF를 0, 10, 50 및 100ng/$m\ell$ 첨가시 상실배기이상 발육된 체외 발육 성적은 각각 19.7, 29.0, 26.6 및 40.4%로서 EGF 100ng1m1 처리구가 여타구보다 통계적으로 유의하게 높은 체외발육 성적을 나타냈으며(P<0.05), PDGF를 0, 1, 5 및 10ng/$m\ell$ 첨가하여 배양한 경우 상실배기 이상 발육된 체외발육성적은 각각 22.0, 21.6, 33.9 및 15.9%로서 PDGF 5ng/$m\ell$ 처리구가 여타구에 비해 높은 체외발육성적을 얻었다. 본 실험에서 체외수정후 생산된 배반포기 수정란의 세포수는 처리구간 커다란 차이는 인정되지 않았다.

The present study was conducted to investigate the effects of antioxidants and growth factors on early development of porcine follicular oocytes fertilized in vitro. The embryos without cumulus cells were transferred into NCSU 23 medium and cultured for 5~6 days. The proportions of embryos developed to morula and blastocyst stages were significantly higher (P<0.05) in medium with 1.0(21.1%) and 2.0mM(22.3%) NAC than in control (14.7%) and 0.5mM NAC (12.2%). When the embryos were cultured in medium with ebselen, the highest proportion(36.9%) of embryos developed to morula or blastocyst stages was obtained in medium with 10 $\mu$M ebselen. Higher proportions of embryos were developed to rnorula and blastocyst stages during culture with 100$\mu$M than control, 50 and 200$\mu$M glutathione. The EGF and PDGF were evaluated as growth factors supplemented to the culture medium for in vitro development of porcine embryos. The proportions of embryos developed to morular and blastocyst stages were significantly higher (P<0.05) in medium with 100ng/m1 ECF than control, 10 and 50ng/$m\ell$ EGF. However, the proportions of embryos developed to morular and blastocyst stages were not significantly different among concentrations of PDGF. In conclusions, these results indicate that NAC, ebselen and glutathione or EGF and PDGF can increase the proportion of embryos that develop beyond morula stage.

키워드

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