본 연구는 돼지 수정란의 동결에 있어서 vitrification 동결 융해 후 내동제의 종류완 농도, PVP 및 sucrse와 trehalose의 첨가가 생존율에 미치는 영향을 조사하고자 수행하였다. 1 Vitrification동결에 이용된 각 발생단계의 체외수정란은 1,063개 중 2세포기는 245$(23.0\%)$개, 배반포는 $256(24.1\%)$, 초기 배반포는 $234 (22.0\%)$, 확장 배반포는 221개 $(20.8\%)$, hatching 배반포는 107개 $(10.1\%)$이었다. 상실배, 초기 배반포 및 확장배반포를 EDS와 ETS로 희석 후 vitrification동결 융해했을 때 생존율은 각각 $69.1\%,;70.3\%,;69.8\%$ 및 $62.5\%,;61.7\%,;63.6\%$로서 EDS군에서 확장 배반포군에서 가장 높은 생존율을 나타냈다. 2. 각 발생단계의 수정란을 vitrification동결 융해했을 때 생존율은 초기 배반포는 $61.1\%$, 확장 배 반포는 $27.8\%$, hatching 배 반포는 $16.7\%$로서 대조군의 $92.3\%,;71.2\%,;55.8\%$에 비해 낮았지만 높은 생존율을 나타냈다. 3. 수정란을 EDS와 EDT내동제에 $10\%$ 및 $20\%$ PVP 액을 첨가하석 희석 후 vitrification 동결 융해했을 때 정상적 발생을 나타내는 수정란은 $74.3\%,;77.5\%$ 및 $79.4\%$ 및 $71.1\%$였다. 동결 융해한 수정란을 $24~48$시간 배양했을 때 $37.1\%,;40.0\%$ 및 $35.3\%,;31.6\%$로서 생존율이 현저하게 감소하였다. 수정란에 EDS와 EDT와 $10\%$ 및 $20\%$의 PVP를 첨가한 내동제를 이용하여 동결 응해했을 때 PVP농도간의 생존율은 유의한 차이가 없었다. 4. 각 발생단계의 수정란을 EDS 내동제로 vitrification 동결 융해 후 배양했을 때 발생율은 상실배는 $58.2\%,;36.4\%,;14.5\%$였고, 초기 배반포는 $62.5\%,;45.8\%,;20.8\%$였고, 확장 배반포는 $74.1\%,;61.1\%,;29.6\%$였고, hatching 배반포는 $60.0\%,;40.0\%,;14.0\%$였다.
The present study examines the effects of kinds and concentrations of cryoprotectants, PVP and sucrose and trehalose on the survival rate of vitrified porcine embryos. 1. The developmental stages for the embryos used in vitrification were $245(23.0\%)$ for 2 cell stage, $256(24.1\%)$ for the blastocyst, $234(22.0\%)$ for the early blastocyst $221(20.8\%)$ from the expanded blastocyst and $107(10.1\%)$ from hatching blastocyst out of 1,063 embryos. 2 The survival rate of morula, early blastocyst and expanded blastocyst vitrification-thawed with EDT and EGS were $69.1\%,;70.3\%,;69.8\%;and;62.5\%,;61.7\%,;63.6\%$, respectively. The expanded blastocyst treated with EDS showed the highest survival rate compared with the other cryoprotectants. 3. The survival rate of early blastocyst, expanded blastocyst and hatching blastocyst vitrification-thawed with EDS diluted in $medium + 10\%$ FCS were $61.1\%,;27.8\%,;16.7\%$, respectively. This result were love. than the control of group $(92.3\%,;71.2\%,; 55.8\%)$. 4. The survival rate of embryos vitrified with EDS and EDT supplemented with $10\%;and;20\%$ PVP were $74.3\%,;77.5\%;and;79.4\%,;71.1\%$, respectively. The survival rate of vitrified embryos cultured for $24~48$ hours were $37.1\%,; 40.0\%;and;35.3\%,;31.6\%$ which were significantly lower than that of non-cultured embryos. The survival rate of embryos vitrified with EDS and EDT supplemented between $10\%;or;20\%$ PVP did not have a significant difference. 5. The survival rate of embryos vitrification-thawed with EDS to morula, early blastocyst, expanded blastocyst and hatching blastocyst were $58.2\%,; 36.4\%,;14.5\%$ to morula, $62.5\%,;45.8\%,;20.8\%$ to early blastocyst, $74.1\%,;61.1\%,;29.6\%$ to expanded blastocyst and $60.0\%,;40.0\%,;14.0\%$ to hatching blastocyst.
