The brilliant cresyl blue (BCB) has been used to select the developmental competent oocytes in pigs, goats and cows. Growing oocytes have a higher level of active glucose-6-phosphate dehydrogenase(G6PDH) compare to mature oocytes and are rarely stained compared to mature oocytes, because G6PDH converts BCB to colorless. First polar body extrusion regard as a guideline of meoisis completion. Selection of polar body extrude oocyte is more developmental competent to blastocyst than unselected. This study was conducted to compare the BCB test to the polar body extrusion on selection of developmental competent porcine oocytes for the production of blastocyst. Cumulus-Oocytes complex were exposed to 26uM BCB stain diluted in NCSU-23 for 90 min. There was no significant difference embryo development to blastocysts between BCB treated and not treated(19.58±1.99 vs 18.75±2.27 %), which means there was no detrimental effect of BCB exposure to oocytes. Normal fertilization is not differed among treatment groups from 70.0 to 78.4% development to blastocyst, beside polyspermy did not. To compare two different selection methods, BCB test and polar body extrusion, evaluate the developmental competent of IVP embryos. BCB+PB+(blue stained and polar body extruded, 20.71±0.45%) and BCB－PB+(colorless and polar body extruded, 20.04±1.29%) groups are significantly (p<0.05) higher developed than those of BCB+PB－(blue stained and no polar body, 13.24±0.73%) and BCB－PB－(colorless and no poladbody, 7.25±0.77%). These results showed that selection of polar body extruded oocytes method is more efficient than that of BCB test.

본 연구에서 돼지 난포란에서 채취된 난모 세포들을 체외성숙 후 형태적으로 선별하거나 극체 방출란을 선별하여 활성화 처리 후 48시간째에 분할란을 선별할 때 배발달율이 어느정도 향상되는지를 검토하였다. 난모 세포를 48시간 성숙 배양 후 형태적 선별과 극체의 방출 유무를 검사하고, 선별된 난모 세포들을 시간 추가 배양한 후 7% ethanol로 활성화시키고 cytochalasin B에 5시간 노출 후 PZM-5 배 양액으로 7일간 배양하였으며, 배양 중

This study was carried out to elucidate whether sperm contain a factor inducing second polar body extrusion and to search for an effective collection method of the sperm factor Thus, sperm extract, dialyzed sperm-extract or liquid chromatographic fractions of sperm extract was microinjected into ovulated oocytes. And the microinjected oocytes were incubated for 24 hours to investigate about the extrusion of second polar body. The results obtained were as follows; 1. Sperm extract significantly increased the second polar body extrusion. 2. Sperm extract showed five major fractions at retention volumes (RVs) 1.25, 1.37, 1.84, 2.10 and 2.67ml after separation with Superose 12 column. These sperm extract fractions did not significantly increase the second polar body extrusion. 3. Dialyzed sperm-extract significantly increased the second polar body extrusion 4. Dialyzed sperm-extract showed three maior fractions at RVs 1.88, 2.14 and 2.77ml after separation with Superose 12 column. Of these fractions, the fraction RV2.14 significantly increased the second polar body extrusion. In conclusion, sperm extract contained a factor inducing the second polar body extrusion and the factor was contained largely in fraction RV2.14 after dialysis and liquid chromatographic fractionation of sperm extract.