• Development and Reproduction
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• v.10 no.4
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• pp.239-245
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• 2006

Experiments were conducted to determine the effects of beta-mercaptoethanol(${\beta}-ME$) supplements to the maturation medium on in vitro fertilization(IVF) and intracellular glutathione(GSH) concentration. Bovine cumulus-intact oocytes were matured in TCM-199 medium containing FBS, hormonal supplements, and ${\beta}-ME$(0, 25 and $50\;{\mu}M$) for 12h and 24 h. After culture, cumulus-free matured oocytes were co-incubated with frozen-thawed spermatozoa for 24h. Maturation rate increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant differences among treatment groups. Also, increases(p<0.05) in intracellular GSH concentration before and after fertilization were observed in $50\;{\mu}M\;{\beta}-ME$ supplements to the maturation medium. Male pronuclear formations after IVF was increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant difference among treatment groups. In conclusion, supplementing ${\beta}-ME$ into the maturation medium increased maturation rates, fertilization rates, and intracellular GSH concentrations.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.133-142
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• 2002

The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.

• Development and Reproduction
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• v.3 no.1
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• pp.21-28
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• 1999

In order to examine the effect of $Ca^{2+}$ -chelation on in vitro fertilization of rat zona-free oocyte, the formation of cortical granule envelope (CGE) and the rate of fertilization related to monospermy and polyspermy were determined. The ultrastructural characteristics of oocytes were observed with the scanning electron microscope and BAPTA/AM was used for calcium-chelation. The CGE formed by cortical reaction was observed in zona-free oocyte inseminated in vitro and it was also observed in the calcium chelator (1, 5, 10$\mu$M BAPTA/AM) treated zona-free oocytes inseminated in vitro. The CGE developed according to incubation time. The fertilization rate was decreased in the calcium chelator-treated group (59.8, 38.1, 37.0%) compared to the control group (60.6%) but monospermy rate was increased in the calcium chelator-treated group (45.0, 47.3, 50.9%) compared to control group (37.5%). The above results demonstrate that the CGE is formed during fertilization in rat and the extracellular calcium is used in cortical reaction. Also the results suggest that proper concentration of free calcium in oocyte acts as important factor in fertilization.n.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.2
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• pp.87-94
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• 2007

Objective: To examine determinants of successful pregnancy and evaluate COH-IVF outcomes of infertile patients after conservative surgical treatment of borderline ovarian tumor (BOT). Methods: In women of BOT (n=93), from January 1995 to December 1999, 44 of 93 women underwent conservative surgical treatment. From theses 44 women, patients characteristics, surgical and histological parameters were compared between 14 women who conceived and 30 women who failed to conceive. For 5 infertile women of 30 women who failed to conceive, 10 attempt IVF cycles were analysed; clinical pregnancy rate (CPR), implantation rate (IR) and live birth rate (LBR). Results: Women who conceived tend to be younger (25.9 vs 27.0 years) and lower serum CA-125 level (59.7 vs 72.9) compared to women who failed to conceive without significant difference. For 8 cycles out of 10 attempt IVF cycles, except for 2 cancellation cycles, the mean number of oocytes retrieved was 5.6 (range 2$\sim$16) with a mean fertilization rate of 74.4%. The CPR, IR and LBR per embryo transfer were 50.0% (4/8 cycles), 31.6% (6/19) and 50.0% (4/8 cycles) respectively. During the mean follow-up period after COH-IVF initiation, 29.6 (range 14$\sim$61) months, no recurrence was found. Conclusion: No determinant of successful pregnancy was found after conservative treatment for BOT. COH-IVF may be considered for infertile patients after conservative treatment of BOT. However, larger clinical studies with longer follow-up are necessary to evaluate the safety and efficacy of COH-IVF. All patients should be informed of the potential risks associated with ovarian hyperstimulation and close follow-up is necessary after COH-IVF.

• Journal of Embryo Transfer
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• v.9 no.1
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• pp.53-63
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• 1994

소 체외수정란의 체외 발육에 $CO_2$와 $O_2$의 농도가 미치는 영향에 대하여 두가지 배양액과 서로 다른 Co-culture 체계를 이용하여 검토하였다. 체외 수정후 40~44시간에 회수간 2~8 세포기 수정란을 여러가지 gas조건하의 다른 배양조건에서 무작위로 옮겨 실험을 수행하였다. 5% $CO_2$와 $O_2$,10% $O_2$및 20% $O_2$조건에서 배양을 실시한 결과, 상실배가 이상 발육된 체외 발육성적은 각각 22.1% 15.0% 및 21.1%였다.(p<0.05). 한편 배양액에 따른 헤외 배양성적은 KOSM배양액(19.1%)의 경우가 Menezo's B2 배양액(13.7%)에서 배양한 경우보다 더 높은 성적을 얻었다.(p>0.05,Table 1). 2~8세포기 수정란을 5% $CO_2$또는 10% $CO_2$와 5%, 10% 및 20%의 $O_2$조건 하에서 체외 배양을 실시한 경우 각각 15% 와 8%의 체외 발육 성적을 얻었고(P>0.05), $O_2$조건의 경우 10% $O_2$(17%)와 20% $O_2$(20%)의 배양조건을 이 5% $O_2$(26%)보다 낮은 성적을 나타냈다. (P<0.05), (Table2). 체외 수정 후 얻은 2~8세포기 수정란을 단순 배양액 과 두개의 다른 공동 배양체계를 이용하여 5% $CO_2$와 5% 및 20% $O_2$의 배양 조건하에서 체외 배양을 실시한 결과 상실배와 배반포기 까지 발육된 체외 발육 성적은 배양액과 공동배양 체계에 따른 차이는 인정되지 않았다(p>0.05). 그러나 $O_2$의 농도는 소 체외 수정란의 체외 발육성적에 영향을 미치는 것으로 나타났다.(Table 3). 본실험은 $CO_2$와 $O_2$의 농도가 소 체외수정란의 체외 배양할때 배양체계에 따라 다른 영향을 미치는 것으로 나타났다.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.275-282
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• 2004

This study was carried out to examine the effects of development media and those change on in vitro development (IVD) of porcine oocytes matured and fertilized in vitro. Putative embryos, which were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10％ porcine follicular fluid (pFF) and were fertilized in mTBM, were developed in vitro as the experimental scheme. The results are as follows. When porcine putative embryos were cultured in vitro in NCSU-23 or CZB/Pig-MEM, the percentage of oocytes cleaved was not different between two systems, but the percentage of blastocysts in NCSU-23 was significantly higher than in CZB/Pig-MEM (P<0.05). And when porcine putative embryos were cultured in vitro in NCSU-23 during 7 days with or without changing media at day 5, which was supplement with or without 10％ fetal bovine serum(FBS), the percentage of oocytes cleaved, blastocysts at day 6, and the cell number of ICM, TE and total of blastocysts at day 7 were not different among three treatments. As a result of this study, it is supposed that NCSU-23 be more favorable, to develop porcine embryos derived from IVM/IVF, than CZB/Pig-MEM, but that demonstration on the effects of changing medium with fresh one stand in need of the more experiments.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.116-116
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• 2003

도축 소의 난소를 이용하여 체외에서 수정란 생산과 이식이 산업화에 접어들고 있지만, 그 기원이 되는 난소의 자질은 검토되어 있지 않고, 생산된 송아지의 자질 또한 의문시 되고 있는 실정이다. 본 실험에서는 도축 한우의 육질과 육량에 따른 배발달율을 조사하여 고품질 체외수정란의 생산에 기초를 확립 하고자 실시하였다. 한우 난소는 도축 직후에 개체별로 paper에 싸서, 0.9%생리식염수 (25-$28}{\circ}C$)가 들어있는 보온병에 담아 실험실로 운반하였다. 운반된 난소의 2~8mm의 가시난포로부터 난포란을 회수하였다. 회수된 난포란은 10% FBS, 1$\mu g/ml$ FSH, 10$\mu g/ml$LH 그리고 1$\mu g/ml$ Estradiol-$17 \beta$가 첨가된 TCM199 용액에서 24시간 체외성숙을 실시하였다. 체외수정은 fer-TALP 용액을, 체외배양은 CR1aa 용액에서 배양 3일째까지는 0.3% BSA, 그 이후에는 10% FBS와 난관 상피세포를 첨가하여 사용하였다. 통계분석은 $X^2-test를 이용하였다. 도축 한우의 육질등급에 따른 수정율은 1, 2, 3 및 등외등급에서 각각 63.7, 82.7, 73.2 및 84.0%로서 등외등급에서 가장 높은 수정율을 나타냈다. 배반포까지 발달율도 각각 17.1, 32.2, 26.8 및 40.0%로서 등외등급에서 가장 높았으며 특히 등외등급의 배발달율이 1등급에 비하여 유의적(P<0.05)으로 높았다. 육량등급에 따른 수정율은 A, B, C 및 등외등급에서 각각 90.0, 62 0, 69.2 및 85.0%로서 A등급이 가장 높았고 배반포까지 발달율은 각각 21.2, 18.7, 22.5 및 20.2%로서 C등급이 가장 높았으나 유의적인 차이는 없었다. 이상의 결과에서 한우 난포란의 배발달에는 육량등급보다는 육질등급에 많은 영향을 받는 것으로 판단된다. 한편 육질 1등급에서 배발달율이 낮은 이유는 육질 향상을 목적으로 암소를 비육 하는 경우 발생하는 번식장애와 밀접한 관계가 있는 것으로 사료된다.

• 대한생식의학회:학술대회논문집
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• 1995.05a
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• pp.8-11
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• 1995

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.58-58
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• 2003

포유동물 체외 수정란의 체외 발육율을 향상시키기 위해서는 배양액내에 생성되는 free radical을 제거하는 것이 한가지 요인으로 지적되고 있다. 체외 배양액내에 생성되는 free radical을 제거하기 위하여 여러 종류의 항산화제 첨가 배양이 많이 이용되어 체외발육성적을 향상시키고 있다. 포유동물의 번식에 있어서 Melatonin은 광주기 조절 작용에 기여하는 호르몬으로서 혈중 Melatonin의 농도는 번식 작용에 중요한 역할을 하며, 또한 수정란의 세포분열과 세포주기에 영향을 미치며 배양액내의 free radical을 제거하여 수정과 초기 수정란 발달에 영향을 미쳐 항산화 기능이 있는 것으로 보고되고 있다.