본 연구는 인간 난관액 또는 자궁액 내에 존재하는 에너지원이 생쥐 2-세포기 배의 체외 발달에 미치는 영향을 조사하기 위하여 실시하였다. ICR 암 생쥐에 5 IU hCG 주사 후 46～50시간에 2-세포기 배를 회수하였다. 회수된 배는 3가지 배양 조건 [대조군: 0 mM, Group A: glucose(G) 0.5 mM + pyruvate(P) 0.32 mM + lactate(L) 10.5 mM, Group B: G 3.15 mM + P 0.1 mM + L 5.83 mM]에서 72시간 배양하였다. 배양 24 시간에 상실배 출현율은 group A (72.3%)와 group B (56.6%)가 대조군(34.9%)보다 유의하게 높았다(p<0.05). 그러나 48시간에 배반포기 배 출현율은 대조군(51.8%)이 group A (39.8%)와 group B (28.9%)보다 유의하게 (p<0.05) 높았다. 72시간에 투명대 부착 (ZiB, 41.0～51.8%), 투명대 탈출 (ZeB, 18.1～32.5%) 및 총 배반포기 배 출현율 (68.7～73.5%)은 실험군 간에 통계적인 차이가 없었다. 배반포기 배의 평균 세포수와 ICM 세포수는 group A (70.8, 13.4)와 group B (64.4, 11.8)가 대조군 (53.1, 5.7)보다 유의하게(p<0.05) 많았고, 통계적인 유의차는 없었으나 group A가 group B보다 많은 경향이었다. 총 세포수에 대한 ICM 비율은 group A(22.9%)와 group B(23.7%)가 대조군(14.2%)보다 유의하게(p<0.05) 높았다. 영양배엽(TE) 세포수(34.1～45.1)는 실험군 간에 통계적인 차이가 없었다. ICM에 대한 TE 비율(ICM:TE ratio)은 대조군(1:6.0)이 group A(1:3.4)나 group B(1:3.4)보다 유의하게(p<0.05) 높았다. 생쥐 2-세포기 배를 배양하여 72시간까지의 배 발달율을 살펴보면 배양액에 에너지원을 첨가하는 것이 효과적이었으며, 자궁액 농도보다는 난관액 농도로 에너지원을 조절했을 때 배 발생 능력이 높은 경향을 보였다.

The efficient cryopreservation of embryos requires optimal times of dehydration and rehydration This study was carried out to investigate the effect of various times of dehydration and rehydration The effects were evaluated through testing morphological normality and developmental ability of 1 cell mouse embryos which were ultrarapidly frozen and thawed. The 1 cell embryos were dehydrated for 1.5, 3, 5, and 10 minutes using mPBS-BSA containing 3.SM DMSO and 0.25M sucrose on cooling chamber or on ice. After ultrarapidly frozen and thawed, they were rehydrated for 0, 0.5 and 5 minutes with mPBS-BSA containing 0.25M sucrose at room temperature. The results obtained were as follows: The embryos that were rehydrated for 0.5 minutes showed higher normality than the embryos for 0 and 5 minutes did. The embryos that were dehydrated for 10 minutes showed higher normality than the embryos for 1.5, 3, and 5 minutes did. The developmental ability of normal thawed-embryos was high in 10 minute dehydration treatment compared to other treatments. However, it was not affected by cooling methods (on ice and on cooling chamber) for embryo dehydration.

This study was carried out to efficiently use the ultrarapid freezing method in the cryopreservation of mouse ova. For this, the effects of dehydration method, oval vigour and controlling method on post-thawing viability were investigated. Fresh mouse ova were dehydrated in mPBS with 3.5M DMSO and /or 0.25M sucrose, and directly immersed in L for ultrarapidly freezing. The frozen ova were thawed at 37, rehydrated in mPBS with 0.25M sucrose, and then repeatedly washed in HAM's Fl0 before evaluating the morphological normality of frozen-thawed ova. The results obtained showed that there was difference between treatments in a experiment. 1) The post-thawing viability of ova dehydrated in multi-step (48.413.8%) was higher than that of ova in two-step (40.914.0%). 2) The post-thawing viability of fertilized ova (8714.0%) was significantly(p<0.0l) higher than that of unfertilized ova (5.45.4%). 3) The post-thawing viability of ova dehydrated and rehydrated using a cooling machine (95.84.2%) was significantly(p<0.05) higher than that on ice(84.19.9). In conclusion, in order to efficiently cryopreserve ova in vitro with ultrarapidly freezing method, highly viable embryos should be selected, heavy osmotic shock to the dehydrating ova should be avoided, and embryos in high osmotic condition were dehydrated and rehydrated in a constantly low temperature.

본 연구는 자궁근종 성장에 관한 분자생물학적인 기전의 이해를 위해 자궁근종 및 정상 자궁근세포의 초기 배양방법을 확립하기 위해 실시하였다. 이를 위해 최종적으로 두 가지 세포 배양 방법이 확립되었다. 그리고 안정적으로 연구(특히, 여성호르몬에 대한 반응 연구)에 사용할 수 있는 가장 적합한 세포 배양 방법이 모색되었다. 두 가지 세포 배양 조건 중 두 번째 방법(method 2)이 안정적으로 세포의 반응을 연구하는데 더 나은 방법으로 결론 내려졌고, 여