• Clinical and Experimental Reproductive Medicine
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• v.37 no.4
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• pp.329-338
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• 2010

Objective: This study was carried out to know whether cryopreservation of sibling 2PN zygotes could increase the cumulative delivery rates in the patients who had less than 10 fertilized zygotes. Methods: A retrospective analysis was performed in 138 in vitro fertilization-embryo transfer (IVF-ET) cycles with less than 10 fertilized zygotes during January 2003 to December 2007 in Cheil General Hospital. These cycles were divided into two groups. In Group I (n=86), all fertilized embryos were cultured to transfer on day 3 without cryopreserved embryos at the 2PN stage. In Group II (n=52), among fertilized zygotes, some sibling zygotes were frozen at the 2PN stage, the remainder were cultured to transfer. Clinical outcomes in fresh ET cycles and cumulative ongoing pregnancy rates after subsequent frozen-thawed (FT)-ET cycles were compared. Results: There were no significant differences in female mean age, number of retrieved oocytes and total fertilized embryos between two groups, Number of cultured embryos was significantly lower in Group II ($5.2{\pm}0.5$) than in Group I ($8.4{\pm}0.7$) (p<0.01). Also, number of transferred embryos was significantly lower in Group II ($3.3{\pm}0.6$) compared with Group I ($3.6{\pm}0.6$) (p<0.01). ${\beta}$-hCG positive rates and delivery rates (51.2 vs. 46.2 % and 41.9 vs. 34.6 %, respectively) after fresh ET were slightly higher in Group I than in Group II. However, the differences were not statistically significant. Also, the cumulative delivery rates after subsequent FT-ET cycles were not significantly different between Group I (48.8%) and Group II (50.0%). Conclusion: This study showed that cryopreservation of sibling 2PN zygotes from patients who had less than 10 zygotes in the fresh ET cycles did not increase cumulative delivery outcomes. But, it could provide an alternative choice for patients due to offering more chance for embryo transfers if pregnancy was failed in fresh IVF-ET cycles.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.247-253
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• 2006

본 연구는 안정된 돼지 체외 성숙 난자를 얻을 목적으로 배양액의 종류 및 배양액에 EGF, ${\beta}-ME$, 호르몬 첨가가 돼지 난포란의 체외 성숙에 미치는 영향을 조사하였다. 난포란을 TCM-199, NCSU-23 및 PZM-3으로 48시간 배양했을 때 체외 성숙율은 각각 $22.1{\pm}0.70%,\;30.6{\pm}0.70%$ 및 $30.4{\pm}2.82%$였다. TCM-199로 48시간 배양했을 때 체외 성숙율은 NCSU-23 및 PZM-3 보다 약간 낮은 체외 발생율을 나타냈다. 난포란을 25 ng/ml의 EGF를 첨가한 TCM-199, NCSU-23 및 PZM-3로 48시간 배양했을 때 체외 성숙율은 각각 $46.3{\pm}2.8%,\;76.6{\pm}3.1%$ 및 $72.2{\pm}2.6%$로 나타났다. 난포란의 배양 시 배양액에 25 및 50 ng/ml의 EGF를 첨가 후 48시간 배양했을 때 첨가하지 않은 군에 비해 높은 체외 성숙율을 나타냈다(p<0.05). 난포란을 NCSU-23 및 PZM-3 배양액에 $25{\mu}M/ml$의 ${\beta}-ME$를 첨가한 후 48시간 배양했을 때 체외성숙율은 각각 $43.9{\pm}1.41%,\;41.7{\pm}l.41%,\;44.4{\pm}0.70%,\;40.6{\pm}0.70%$로 나타났다. 난포란을 $25{\mu}M/ml$의 ${\beta}-ME$를 첨가한 NCSU-23로 48시간 배양했을 때 첨가하지 않은 군에 비해 높은 체외 성숙율을 나타냈다(p<0.05). 난포란의 배양 시 NCSU-23에 PMSG, hCG, PMSG+hCG, hCG+${\beta}$-estradiol, PMSG+${\beta}$-estradiol을 첨가 후 배양하였을 때 체외 성숙율은 각각 75.6%, 77.8%, 80.0%, 86.4% 및 84.8%로서 무첨가 군(64.4%)에 비해 높게 나타났다(p<0.05).

• Journal of Embryo Transfer
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• v.5 no.1
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• pp.21-27
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• 1990

The technique for maturation of follicular oocyte has been devised to provide such a low cost and in ptentifut number supply of bovine embryo. Some of problems concerning production of bovine embtyo in vitro were discussed in this paper. Bovine follicular oocytes cultured in vitro achieved normal fertilization but cleavage rates to blastocyst were low compared to the oocyte matured in vivo. It has been concluded that a deficient cytoplasmic maturation occurs in the oocytes matured in vitro. These results indicate that the studies for maturation of bovine follicular oocytes in vitro need improvement of culture conditions and to define the characteristics that might be indicative of healthy oocyte.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2005.10a
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• pp.93-93
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• 2005

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.141-141
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• 2003

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.109-109
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• 2004

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 1997.10a
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• pp.48-48
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• 1997

• Journal of Embryo Transfer
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• v.10 no.2
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• pp.139-144
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• 1995

10% ethanol에 의한 처녀발생유가 및 체외수정된 돼지 난포란을 CZB와 CRlaa 에서 배양하여 배발달율을 조사하였다. 또한 CZB에 각기 다른 농도의 cholesterol (0$\mu$g/mL, 2$\mu$g/mL, 5$\mu$g/mL, 10$\mu$g/mL)을 첨가한 후 체외수정된 돼지 난포란을 배양하여 배발달률을 조사하였다. CZB 구는 BOEC와 공배양하였다. 처녀발생유가 48시간 후 2~8세포기로 발달한 난자의 비율은 CZB 구가 32.2%, CRlaa구가 16.8%였으며, 192시간후 상실배로 발달한 난자의 비율은 CZB구가 5.75%, CRlaa구가 0%였다. 체외수정 48시간 후 2~8세포기로 발달한 난자의 비율은 CZB 구가 42.4%, 81%, CRlaa 구가 17.6%였으며, 192시간 후 상실배로 발달한 난자의 비율은 CZB 구가 5.81%, CRlaa 구가 0%였다. 각기 다른 농도의 cholesterol이 첨가된 CZB구에서 상실배로 발달한 난자의 비율은 각각 0,10,0.45,0%였다.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.137-146
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• 2006

The objective of this study was to examine the effect of donor cell types, the source of recipient oocytes and estrous synchronization on pregnancy and delivery rates of somatic cell nuclear transfer (SCNT) embryos in Korean native goats. Recipient oocytes were surgically collected after superovulation. Ear cells and fetal fibroblasts were collected and cultured in serum-starvation condition (TCM-199 + 0.5% FBS) for cell confluence. The zonae pellucidae of in vivo- and in vitro-matured oocytes were partially drilled using a laser system. Single somatic cell was transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3 M mannitol. After the fusion, embryos were activated by Ionomycin+6-DMAP. NT embryos were cultured in mSOF medium supplemented with 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 12 to 20 hr. One hundred and two SCNT embryos were transferred into 20 recipients and pregnancy rate at days 30 was 20.0%. Of them, one developed to term and delivered 1 kid. Ear cells showed significantly higher fusion (63.8 vs. 26.5%) and pregnancy rates (20.0 vs. 0.0%) than those of fetal fibroblast (p<0.05). The recipients synchronized by CIDR showed significantly lower pregnancy rates compared to that of recipient in natural estrus ($0.0{\sim}25.0%$ vs. 100%) (p<0.05). Cloned kid was born from the recipient in natural estrus. For the synchronization of estrus between recipient and donor, there was no difference between treatments (${\pm}0$ vs. +12 hr) in pregnancy rate. The first healthy cloned kid (Jinsoonny) was produced by transfer of SCNT embryos derived from in vivo oocytes and ear cells into a recipient goat whose estrus was synchronized with the donor. These results imply that donor cells for nuclear transfer may affect the success rate, and the estrus synchronization between donor and recipient animals can also be important.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.107-110
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• 2007

본 연구는 BPA 및 nicotine 첨가 농도와 배양 시간이 돼지 난자의 체외성숙에 미치는 영향을 조사하였다. $0.02{\sim}10.0mM$ BPA와 $0.5{\sim}10.0mM$ nicotine이 첨가된 TCM-199배양액에서 $40{\sim}52$시간 난자를 배양했을 때 체외성 숙율을 조사하였다. BPA농도가 높을수록 체외성숙율이 유의적으로 낮게 나타났다. $0.05{\sim}10.0nM$ BPA를 첨가한 TCM-199 배양액에서 난자를 44시간 배양했을 때 체외성숙율은 각각 $40.0{\pm}4.1%,\;24.0{\pm}4.7%,\;10.0{\pm}5.3%,\;6.0{\pm}3.2%,\;0.0{\pm}0.0%$로서 첨가 농도가 증가할수록 낮은 체외성숙율을 나타냈다. 난자를 $0.5{\pm}10.0mM$ nicotine를 첨가한 TCM-199 배양액에서 44시간 배양했을 때 체외성숙율은 각각 $44.0{\pm}4.5%,\;24.0{\pm}4.2%,\;18.0{\pm}4.9%,\;8.0{\pm}2.2%,\;0.0{\pm}0.0%$로서 대조군$(52.0{\pm}4.5%)$에 비해 낮은 체외성숙율을 나타냈다. 난자를 0.5 nM BPA와 2.5 mM nicotine을 첨가한 TCM-199에서 $40{\sim}52$시간 배양했을 때 체외성숙율은 $8.3{\pm}2.1%{\sim}26.0{\pm}3.9%$ 및 $11.2{\pm}2.2%{\sim}28.6{\pm}3.9%$로서, 44시간 배양이 다른 배양시간보다 가장 높은 체외성숙율을 나타냈다.