This study was performed to investigate the effect of types of oil (OVOIL vs. OIL) on B6D2F1 mice oogenesis. In this study, B6D2F1 F1 mice were used in order to maximize oogenesis. The expansion rate of cumulus cells (82.0%±0.2 vs. 78.0%±0.1), in vitro fertilization rate (92.0%±0.1 vs. 88.0%±0.1), developmental rate (91.0%±0.1 vs. 87.0%±0.2), blastocysts formation rate (56.0%±0.1 vs. 57.0%±0.1), and zona hatched rate(41.4%±0.2 vs. 24.0%±0.1) were not different between groups (NS; P>0.05). However, there was a significant difference in maturation rate; the OVOIL group showed higher maturation rate compared to that of the OIL group (96.0%±0.1 vs. 87.0%±0.1; P<0.05). In the blastocysts cell numbers, the total cell numbers (83.9±26.1 vs. 56.9±23.9), ICM cell numbers (15.7±8.8 vs. 6.3±3.5), TE cell numbers (68.3±25.7 vs. 50.7±24.1), % ICM (21.6%±0.1 vs. 12.7%±0.1), and the ratio of ICM:TE (1:6.2±6.5 vs. 1:10.3±7.0) were significantly higher in the OVOIL group than the OIL group (P<0.05). These results suggested that it is expected to achieve the more developmental ability of B6D2F1 mice depending on the type of oil (OVOIL vs. OIL). In addition, the results can provide essential information for culture condition on B6D2F1 mice. Henceforth, thus, it is expected that these results herein might be used for in vitro culture of human embryos.

This study was conducted to evaluate the effect of transfer temperature of epididymis on survival rate of semen and development ability of B6D2F1 mice embryos. No significant differences were noted in the survival rate of semen (59.0% ± 0.1 vs. 47.6% ± 0.1), in vitro fertilization rate (90.7% ± 0.1 vs. 90.7% ± 0.1), developmental rate (90.0% ± 0.1 vs. 90.0% ± 0.1), and blastocysts formation rate (53.1% ± 0.2 vs. 52.3% ± 0.2) between groups. (NS; P>0.05). However, the zona hatched rate was significantly higher in the 4°C group compared to those of the 37°C group (47.8% ± 0.1 vs. 25.6% ± 0.2; p<0.05). When it comes to cell numbers of blastocysts, the % ICM (/total cells) was significantly higher in the group of 4°C compared to the 37°C (27.0% ± 0.1 vs. 18.3% ± 0.1; p<0.05). However there were no differences in total cell numbers (72.7 ± 31.6 vs. 62.0 ± 36.6), ICM cell numbers (17.0 ± 7.8 vs. 14.6 ± 8.6), TE cell numbers (55.8 ± 29.8 vs. 64.0 ± 24.4), the ratio of ICM:TE (1:4.2 ± 4.1 vs. 1:6.4 ± 7.2) between two groups (NS; P>0.05).Taken altogether, it is expected to achieve the best developmental ability of B6D2F1 mice embryos in the transfer temperature of epididymis. Also these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice. In future, therefore, it is expected that results herein might be applied for in vitro culture of human embryos.

This study was conducted to evaluate the effects of different volume (100 μl vs. 2 ml) of microdrop culture on B6D2F1 mice oogenesis. In the present study, B6D2F1/CrljOri F1 mice were utilized in order to maximize oogenesis. Also we used TCM-199, Dulbecco's medified Eagle's medium (DMEM), embryo culture medium (Fertilization medium, Cleavage medium, Blastocyst medium), G series medium and One step medium. Blastulation rate was not different between groups (58.4±2.9% vs. 61.2±4.8%). Zona hatched rate (38±15.4% vs. 27±3.4%) and attached rate (55±13.9% vs. 46±3.9%) did not differ by the volume of culture media. Total cell numbers (59.8±9.7 vs. 70.3±8.7), ICM cell numbers (15.8±0.6 vs. 16.8±1.5), TE cell numbers (44.0±9.7 vs. 53.6±7.3), % ICM (26.4±2.9% vs. 23.8±3.3%) and ICM:TE ratio (1: 2.8±0.4 vs. 1: 3.2±0.6) were not different between groups (i.e., 100 μl vs. 2 ml). These results show that the capacity of the culture medium did not effect the cell numbers of B6D2F1 mice blastocysts. In summary, these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice.

This study was conducted to evaluate the effects of type of culture media (BM, G2, OS, TCM, and MEM) on B6D2F1 mice oogenesis. In the present study, B6D2F1/CrljOri F1 mice were utilized in order to maximize oogenesis. Also we used TCM-199, Dulbecco's medified Eagle's medium (DMEM), embryo culture medium (Fertilization medium, Cleavage medium, Blastocyst medium), G series medium and One step medium. In vitro maturation was highest in BM followed by the order of OS, MEM, TCM and G2 (90±2.8% > 88±3.2% > 85±4.9% > 78±10.2% > 64±7.7%, respectively). To note, the G2 group was statistically different compared to other groups (p<0.05). On the other hand the fertilization rate was highest in the G2 group followed by BM, OS, TCM, and MEM (87±7.2% > 85±6.9% > 74±14.0% > 71±13.8% > 2±1.4%, respectively). The MEM group was significantly lower compared to other groups (p<0.05). The developmental rate was highest in the OS group followed by the G2 group and the BM group albeit no statistical significance was noted (73±11.6% > 71±9.2% > 66±10.4%). Of note, all cells of the TCM and MEM groups were died during embryonic development. The zona hatched rate (51±9.8% vs. 50±9.1% vs. 47±7.2% for BM, G2, and OS respectively) and attached rate (45±12.3% vs. 38±16.1% vs. 37±11.5% for BM, G2, and OS respectively) were not different amongst groups. No difference was found in total cell numbers (74±13.9 vs. 64±9.2 vs. 76±6.7 for BM, G2, and OS respectively), ICM cell numbers (20±1.9 vs. 14±1.8 vs. 15±2.1), TE cell numbers (55±12.5 vs. 49±10.7 vs. 61±5.9), % ICM (30±2.8% vs. 24±7.0% vs. 22.8±2.2%) and ICM:TE ratio (1:2±0.5 vs. 1:3.1±0.8 vs. 1:3.1 ±0.5) amongst groups. In summary, these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice.

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.