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.