• Journal of Embryo Transfer
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• v.19 no.2
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• pp.165-172
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• 2004

This study was carried out to examine the effects of maturation media on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-37 (mNCSU-37), modified NCSU-23 (mNCSU-23), or TCM-199 supplemented with 10% porcine follicular fluid (pFF). Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium(mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU-23. The results are as follows. 1. In the result of IVM, the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different among the media, though numeric value of them were slightly lower in TCM-199 than in mNCSU-37 or in mNCSU-23. 2. In the result of IVF, though the rate of sperm penetration was not significantly different among the maturation media, the percentage of oocytes with male pronucleus (MPN) of ones matured in mNCSU-37 (88.0%) was significantly higher than in TCM-199 (71.1%) (p<0.05). 3. In the result of IVD, the percentage of cleaved oocytes of ones matured in mNCSU-37 (52.3%) or in mNCSU-23 (53.7%) was significantly higher than in TCM-199 (43.1%) (p<0.05), but the rate of blastocysts at day 6 was not significantly different among the maturation media, though putative embryos from oocytes matured in mNCSU-37 or in mNCSU-23 were developed more than in TCM-199. These results suggested that mNCSU-37 or mNCSU-23 was more appropriate than TCM-199 as IVM medium for porcine immature oocytes.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.121-128
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• 2006

In this study, we examined the effects of molecular weight, concentrations and treat the duration of polyvinylpyrrolidone (PVP) in vitro maturation (IVM) medium (Experiment 1), and the effect of PVP in IVM, in vitro fertilization (IVF) and in vitro culture (IVC) medium on the development and cell number of porcine embryos (Experiment 2). The base mediums were NCSU 23 solution for IVM, mTBM solution for IVF and PZM3 solution for IVC. In experiment 1, the development rates to 2 cell and blastocyst stage were not differ from the different molecular weight (MW), concentration and duration of PVP in IVM medium. However, the hatching rate of blastocyst was significantly higher in the group of MW 40,000, 0.5% and $0{\sim}44hr$ than in the other groups (p<0.05). In experiment 2, the results of IVM, IVF and IVC medium with (W) or without (W/O) 0.5% MW 40,000 PVP are follows. The development rate to 2 cell stage was highest in the group of W-W/O-W (p<0.05). The development rate to blastocyst and hatching rate was higher in the group of W-W/O-W and W-W/O-W/O than that of other treatments (p<0.05).

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.19-19
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• 2003

소 난자 체외성숙에 있어 소 태아혈청 (fetal bovine serum: FBS)첨가시 높은 성숙율을 보이며 핵 이식 후 배 발달율에 있어서도 높다. 하지만 소 태아혈청은 hormone, growth factor, vitamine 그리고 다수의 어떤 잘 알려지지 않은 인자를 포함한 복잡한 배지로 알려져 있으며, 수정란 이식 후 태아의 발육중 및 태어난 직후에 발생되는 송아지에서 나타나는 몇몇 비정상적인 현상들의 원인인 것으로 보고되고 있다. 본 실험은 체외 배양시 소 태아혈청과 이의 대체물질로서 BSA 또는 PVA가 첨가된 배양조건에서의 복제 수정란의 배 발달율을 비교함과 동시에 배반포의 세포수 그리고 세포자연사 (Apoptosis)를 각각의 조건에서 비교함으로써 배발달에 미치는 효과를 알아보기 위하여 실시하였다. 핵이식은 소 체세포를 이용하였으며, 핵 이식 후 CR1aa 기본 배양액으로 FBS, BSA, 그리고 PVA를 첨가하여 5% $CO_2$, 5% $O_2$, 39$^{\circ}C$ 조건하에서 7~8일간 배양하였다.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.9
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• pp.510-516
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• 2019

This study examined the effects of fetal bovine serum (FBS), goat blood serum (gBS), and poly-vinyl alcohol (PVA) on the in vitro development and embryo quality of goats for an improvement of embryo production. For the experiment, an in vitro fertilized embryo culture medium was supplemented with 10% FBS, 10% gBS, and 10% PVA to determine their effects on the embryo development efficiency and blastocyst quality. The results showed that the non-serum supplementation group showed significantly lower cleavage rate and blastocyst formation. On the other hand, the gBS and PVA supplementation groups showed a significant increase in the cleavage rate and better blastocyst formation than the control and FBS supplementation group. Furthermore, a TUNEL assay performed to confirm the blastocyst quality showed the same pattern as the embryo development experiment. These results showed that the supplemented gBS or PVA was more efficient in enhancing the in vitro development efficiency of goats than the supplementation of FBS or non-serum. On the other hand, considering the risk of an unidentified factor in gBS, PVA appears to be safer and more efficient in the in vitro development of goat embryos.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.21-26
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• 2007

본 연구는 mSOF(modified synthetic oviduct fluid medium) 배양액을 이용하여 $100{\mu}1$와 $10{\mu}1$ 배양 소적에서 일본 흑우의 수정란 생산 효율을 개선하기 위하여 수행하였다. 난구세포가 부착된 미성숙 난자는 각각 단독 배양조건($S;\;10{\mu}1$ 소적) 및 그룹 배양 조건 ($G;\;100{\mu}1$ 소적)에서 실시하였고 배양액은 TCM-199의 기본 배지에 10% FCS, 0.02IU/ml FSH와 $1{\mu}g/ml$ $estradiol-17{\beta}$를 첨가하여 사용하였다. 배반포 단계로 발육한 수정란은 1.5M ethylene glycol로 직접 이식법에 의한 동결 방법으로 동결을 실시하였고, 세포수는 융해 후 생존 수정란에 대해 조사하였다. 체외 배양 시간이 $16{\sim}17$시간 배양 조건에서 난자의 성숙율은 그룹 배양 조건$(27.1{\pm}16.8%)$보다는 단독 배양조건$(57.1{\pm}15.0%)$에서 성숙율이 높았다(p<0.05). 그러나 체외 배양 시간이 $18{\sim}19$ 시간과 $20{\sim}21$시간 배양시는 유사한 성숙율을 보였다. 난자의 체외 배양율은 체외 배양 시간의 증가에 의해 성숙도가 $86.3{\pm}9.9%$로 증가하였다. 접합체(zygote)의 분할율은 단독이나 그룹 배양 조건에서도 유사한 결과를 얻었다. 배반포 발달율은 배양 $7{\sim}8$일째에 조사한 결과 단독 배양 방법보다는 그룹 배양 방법에서 발달율이 높았으나, 분할된 접합체를 기준으로 한 경우 배반포 발달율$(S;\; 21.4{\pm}10.6%,\;G;\;39.0{\pm}13.1%)$에서는 유의적인 차이가 없었다. 단독 배양과 그룹 배양에서 $6.5{\pm}8$일 사이에 배반포로 발달된 수정란의 세포수 조사에서는 유사한 결과를 얻었다. 동결 융해 후 24시간 배양 후 배반포 생존율(5; 24.2%, G; 30.2%), 부화율(S: 20.9%, G: 12.7%) 및 생존 수정란수(S; 45.2%, G: 42.8%)에서도 배양 조건에 따른 유의적인 차는 없었다. 결론적으로 mSOF배양액을 이용하는 경우 미성숙 난자의 체외 성숙 유도 배양 시 단독이나 그룹 배양 시 배반포 발달율에서 그룹간에 유의적인 차가 인정되었다(p<0.01).

• Korean Journal of Agricultural Science
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• v.23 no.2
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• pp.212-218
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• 1996

This study was conducted to investigate the effects of coculture for the development rate to morula/blastocyst stages of early porcine embryos, derived from oocytes matured and fertilized In vitro, with porcine cumulus cell monolayers(PCM) in the two different media, respectively. The rates of embryos developed to 2-, 4-, 8~16-cell and morula/blastocyst stage were 48.7, 40.5, 34.8 and 17.0% in Ham's F-10 with PCM, and 48.3, 35.6, 22.1, and 13.4% in TCM-HEPES with PCM, respectively. The above development rates to morula/blastocyst stages were significantly higher than those of the embryos cultured in the Ham's F-10 and TCM-HEPES without PCM(P<0.05). The in vitro development rates to the morula/blastocyst stage of I-cell embryos cultured in Ham's F-10 and TCM-HEPES without PCM were 0.0~1.0%. Especially, most of embryos were observed to arrest the development beyond 4-cell stages. As shown in the above results, the coculture of in vitro fertilized porcine embryos with PCM in the two different media enhanced the development of fertilized eggs to morula/blastocyst stages in vitro. However, we didn't find out any difference for the in vitro development to morula/blastocyst stages between Ham's F-10 and TCM-HEPES media.

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.181-185
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• 2004

This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.115-115
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• 2003

• 대한생식의학회:학술대회논문집
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• 2003.12a
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• pp.152-152
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• 2003

• The Journal of the Korean bone and joint tumor society
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• v.11 no.1
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• pp.17-24
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• 2005

Local treatment for tumors has developed from extended radical surgery to function preserving surgery on the basis of modern biology. With the development of minimally invasive technique, it changed to be minimal-invasive surgery. And nowadays technical revolution made non-invasive surgery possible with appearance of several kinds of non-surgical knives such as gamma knife, cyber knife, and HIFU (high intensity focused ultrasound) knife. In this article, history, HIFU machine and treatment procedure, histological change and its mechanism, clinical applications, advantage, disadvantage, and future prospect of extracorporeal high intensity focused ultrasound therapy using HIFU knife will be reviewed.