Effects of Antioxidants on Porcine IVM/IVF Embryos

The purpose of this study was performed to establish the in vitro culture system of porcine in vitro maturation and in vitro fertilization(IVM/IVF) embryo. These studies was to determine the effects of antioxidants(aesculetin, taurine and melatonin) in porcine IVM/IVF embryos. In routine porcine IVM/IVF procedure, oocytes were cultured for 40∼44h incubation in NCSU 23 mediumand matured oocytes were inseminated with frozen semen. Then 2 to 8 cell embryos were removed cumulus cell and were allotted randomly to NCSU 23 containing different concentration of antioxidants in 5% $O_2$ and 5% $CO_2$ at 38.5$^{\circ}C$. Cell numbers of blastocyst were also counted using double fluorescence stain method. Aesculetin were added to NCSU 23 medium at concentration of 1 ug, 5 ug, and 10 ug, when treated with 10 ug(35.7%) of aesucletin at the rate of embryos of the morula plus blatocsyts were higher than those of any other groups (30.2%, 29.5% and 29.2%)(P<0.05). The developmental rates beyond morula stage of porcine embryos in NCSU 23 medium supplemented with taurine 0, 2.5 and 5.0 mM were 26.1%, 26.9% and 31.7%, respectively. The addition of 5.0 mM taurine was higher the developmental rate beyond morula stage than in any other groups. In NCSU 23 medium treated with melatonin 0, 1, 5 and 10 nM, the developmental rate of morula plus blastocysts were 33.3%, 39.1%, 33.3% and 27.9%, respectively. The developmental rate of morula and blascytocys treated with 1nM melatonin was higher than in any other groups(P<0.05). Cell numbers of blastocyst in NCSU 23 treated with melatonin 0, 1, 5 and 10nM were 41.0, 42.6, 39.6 and 33.0, respectively. These results indicate that aesculetin, taurine and melation can increase the developmental rate beyond the morulae and blastocysts in porcine embryos.

돼지 체외수정란의 체외발육에 있어 항산화제의 효과

5% $CO^2와 5% O^2$ 농도 조건 하에서 NCSU23 배양액에 aesculetin을 0, 1, 5 및 10 $\mug/ml$를 첨가한 구에서 상실배기 이상 발육된 체외배양 성적은 10 $\mu$ g 첨가구(35.7%)가 여타구(0 $\mu$g, 30.2% ; 1$\mu$g, 29.5% ; 5$\mu$g, 29.2%)보다 통계적으로 유의하게 높은 성적을 얻었다(P<0.05). NCSU 23 배양액에 taurine을 0, 2.5 및 5.0 mM을 첨가, 체외배양을 실시한 결과 배반포기 이상 발육된 체외발육 성적은 각각 2.8%, 2.2% 및 7.0%였으며, 상실배이상의 체외 발육성적은 26.1%, 26.9% 및 31.7%로서 taurine 5.0 mM 첨가가 체외수정란의 체외발육 성적이 유의적으로 높은 것으로 나타났다(P<0.05). NCSU 23 배앙액에 melatonin을 0, 1, 5 및 10nM을 첨가하여 체외배양을 실시한 결과, 배반포기까지 발육된 체외 발육성적은 17.8%, 26.1%, 20.0% 및 16.3%로서 melatonin 1nM 첨가구가 여타구에 비해 통계적으로 유의하게 높은 성적을 나타냈으며(P<0.05), 상실배기 이상 발육된 체외발육 성적에서는 melatonin 1nM 첨가구가 39.1%로서 대조구 33.3%, 5 nM 첨가구의 33.3% 및 10 nM 첨가구의 27.9%보다 높은 발육율을 나타냈다(P<0.05). 한편 배반포기 수정란의 세포수 조사에서는 melatonin 10 mM 첨가구가 유의하게 낮은 것으로 나타났다.