• Journal of Embryo Transfer
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• v.16 no.2
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• pp.99-106
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• 2001

This study was undertaken to compare the efficiency of recovery rate and development rate of follicular oocytes collected either by aspiration or by slicing method. The follicular oocytes collected by the two methods matured in TCM199 supplemented with 10% steer serum at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. After 22 h of culture, the oocytes were inseminated with frozen-thawed semen (2$\times$10$^{6}$ sperm/ml of final concentration) prepared with Percoll-density gradient in IVF-TALP medium for 16 h. Later, sets of 15 presumptive zygotes were transferred into 50 $\mu$L, droplets of CR1aa medium. On day 4 of the culture, embryos were transferred to TCM199 until day 9. The percentages of nuclear maturation to pre-metaphase II in the oocytes collected by aspiration are significantly (P<0.05) higher than that by slicing (83% vs. 62%, respectively). The mean number of oocytes recovered by slicing per ovary is significantly (P<0.05) higher than that by aspiration (15.1 vs. 6.7, respectively). Although the rates of cleavage and development to blastocyst of oocytes collected b)\\\\`aspiration are significantly (P<0.05) higher than that by slicing, the number of transferable embryos obtained by slicing method is significantly (P<0.05) higher than that by aspiration. From the results. we may conclude that slicing method is better than aspiration method for obtaining large number of transferable embryos per ovary.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.17-21
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• 2015

Alteration in ion channel or transporter expression levels affects cell volume which is produced by movement of water and ion across the plasma membrane. In particular, aquaporin (AQP) channels among ion channels play a crucial role in movement of water across the cell membrane. This study was performed to identify whether AQP expression is changed in bovine follicular cystic follicles using microarray, RT-PCR and Western blotting analyses. In microarray data, AQP4 expression was decreased, whereas AQP7 was increased in cystic follicles. Additional experiments were focused on the AQP7 expression increased in cystic follicles. The microarray data was confirmed by semi-quantitative polymerase chain reaction (PCR) and Western blot analysis. AQP7 mRNA and protein expressions were significantly increased in the cystic follicles (p<0.05). Application of estrogen ($10{\mu}g/ml$) to bovine ovarian cells showed a trend of increase in AQP7 expression. From these results, we suggest that the increase in AQP7 expression in cystic follicles may play an important role in movement of water in bovine ovary. In addition, AQP7, a aquaglyceroporin permeating water and glycerol, could be a good target in development of methods for the cryopreservation of bovine ovary.

• Journal of Embryo Transfer
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• v.21 no.1
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• pp.59-67
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• 2006

To evaluate the functional status of ovarian cyst in Korean native cows, progesterone ($P_4$) and estrogen ($E_2$) level of cystic follicular fluid, ultrasonography for measuring the cystic diameter and thickness of cystic wall, and histological findings were investigated in cystic ovaries from slaughtered Korean native cows. Ovarian cysts were classified as single follicular cyst 51 cows (59.3%), multiple follicular cysts 19 cows (22.1%), single luteal cyst 13 cows (15.1%) and multiple luteal cysts 3 cows (3.5 %) by anatomical and ultrasonography. Ovarian cysts were classified as follicular cysts (54 cows), luteal cyst (16 cows) and non-functional ovarian cyst (16 cows) by hormone analysis, anatomical finding and ultasonography The luteal cyst was accurately diagnosed by cystic wall thickness, but follicular cysts was misdiagnosed 16 cows of 70 cystic cows The cystic fluid $P_4$ concentration was 3.3 ng/ml in follicular cysts and 30.1 ng/ml luteal cysts. There was significantly positive correlations between cystic wall thickness and serum $P_4$ concentration in follicular ($r^2=0.59$, p<0.001) and luteal cysts ($r^2=0.65$, p<0.001). These results indicated that ovarian cysts had various stages of degeneration and luteal cyst was accurately diagnosed measurement of cystic wall thickness by ultrasonography, but follicular cysts were not diagnosed only cystic diameter and cystic wall thickness.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.183-190
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• 2006

These studies were conducted to monitor developmental competence of follicular oocytes collected from the carcass of the high meat quality in Korean native cattle using each individual protocol of IVM, IVF and IVC. The follicular oocytes that were collected from the ovaries of the cow yielded 1, $1^+\;and\;1^{++}$ meat quality were matured, fertilized and cultured using each individual protocol of IVM, IVF and IVC. As results, the number of follicular oocytes collected from individual fundamentally-registered cows yielded 1, $1^+\;and\;1^{++}$ meat grade were 28.9, 28.8 and 29.6 per head, respectively. The rates of blastocyst formation after IVM, IVF and IVC were 27.2, 28.7 and 32.9% in the cows yielded 1, $1^+\;and\;1^{++}$ meat quality, respectively. The rate of blastocyst formation was 8.4 per head. The number of follicular oocytes collected from pedigree registered cows yielded 1, $1^+\;and\;1^{++}$ meat quality were 25.8, 27.1 and 27.0 per head, respectively. The rates of blastocyst formation were 23.0, 33.7 and 42.6% in the meat quality of 1, $1^+\;and\;1^{++}$ after in vitro-manipulation, respectively (p<0.05). The rate of blastocyst formation was 8.5 per head. In conclusion, these results suggest that in vitro embryo production system using individual culture system including IVM, IVF and IVC can make good use of the gene from the carcass of the high meat quality in Korean native cattle.

• Journal of Veterinary Clinics
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• v.7 no.1
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• pp.415-418
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• 1990

To investigate the changes of adrenal gland associated with follicular cyst in Korean native cattle, adrenals ans ovaries were collected from 54 Korean native cattle at abbatoir. Adrenals were stained by H-E stain and they were examined under light microscope. Enlargement of adrenal cortex affecting fascicular and reticular zone appeared in cows associated with follicular cyst.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.37-41
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• 2005

The aim of this study was to improve efficiency and quality of the production of Korean Native Cow embryos. We examined effects of ovarian morphology and maturation vessel on the development and cell number of blastocysts. The development rates to the 2-cell embryos from oocytes collected from the ovaries of different morphological statues were similar ranging between 70.3 and 84.1%. The development rate to the 8 cell- and blastocyst-stage embryos was the highest in the group without both corpus luteum (CL) and follicle. The inner cell mass (ICM), trophectoderm (TE) and total cell number (TCN) were significantly higher in the groups of follicular cyst and regressive CL than other treatment groups, and the same pattern was observed in the ICM/TCN ratio. The development rate to the 2-cell stage was significantly higher in 0.5-㎖ straw group than 0.25-㎖ straw group. However, the development rates to the blastocyst stage were similar between the dish and the straw group. There were no differences in the number of ICM and TE cells, TCN and ICM/TCN ratio of blastocysts from oocytes matured in the different vessels.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.137-141
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• 2007

Antioxidants were well known to be essential supplements in the complex media and serve as a reservoir of oxygen. In this study, Hanwoo COCs (cumulus oocytes complexes) were matured and developed in L-cysteine-TCM199 and analyzed metaphase chromosome. Maturation rate of Hanwoo COCs were 73.4%, 94.6% in 0.1% PVA, 0.1 mM L-cysteine, respectively and showed significantly different between the treatments (p<0.05). Blastocyst formation were revealed 20.3%, 10.0% in 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA, respectively. There were no significant difference among treatment groups. Metaphase chromosome were showed 18.3%, 12.0% in 5% FBS-TCM199, 0.1 mM L-cysteine, respectively and analyzable chromosome were 6.1%, 4.0% and had no differences between the treated groups. In the case of in vitro developmental stages, metaphase chromosome were showed 18.3%, 12.0% in $4{\sim}16$ cells stage, 43.1%, 13.0% in morulae stage and 94.8%, 100.0% in blastocyst stage. These results suggested L-cysteine has beneficial role for in virto maturation and development in Hanwoo COCs.

• Journal of Embryo Transfer
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• v.12 no.2
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• pp.203-209
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• 1997

This study was carried out to compare the actual size(length and height) of ovaries, follicles and corpora lutea of Korean native cow with those on sonograms. We used 3 different probes(3.5 MHz abdominal probe, 6.5 MHz transvaginal probe and 5.0 MHz transrectal probe) and a calipher for measurements of ovaries, follicles and corpora lutea on sonograms and actual size. Under water immersion, 157 ovaries were scanned with 3 probes and measured in actual size and compared each other. The average height and width of ovaries of Korean native cows were 17.40$\pm$3.99 and 34.23$\pm$6.02mm, respectively. In comparison of height, length of ovaries and preovulation follicles, we found that image with a transvaginal probe was nearly the same as the actual size(p<0.01), but with an abdominal probe the image was appeared larger than the actual size. In measurement(diameter) of preovulation follicles the transvaginal probe was proven to be more accurate to the actual size than other probes and in corpus luteum measurement all probes were accurate. In the comparison of number of follicles by different size ranges, there was no statistical difference in the count of follicles over 10 mm in diameter between the transvaginal probe and naked eyes.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.74-74
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• 2001

난자 동결방법의 선별은 보다 효과적인 난자은행의 개발에 필수 불가결한 중요한 요소이다. 이전의 연구에서 마우스의 난자를 ethylene glycol과 electron microscope grid를 이용한 유리화 동결법으로 동결 융해한 결과 기존의 slow freezing 방법에서보다 높은 생존율과 배발달율이 나타남을 관찰하였다. 그러나 동결융해후의 난자는 방추사와 염색체의 이상성이 대조군에 비해 높은 빈도로 나타나 융해후의 배발달율을 감소시키는 것으로 보고되었다. 이에 본 연구에서는 유리화동결법동안 항동해제에 Cytoskeleton system을 안정화시키는 cytoskeleton stabilizer인 taxol을 첨가시킨후 동결시켰을때 생존율과 발달율을 개선시킬 수 있는지 알아보고자 본 실험을 시행하였다. ICR mouse의 성숙란을 채취하여 연구목적에 따라 taxol을 첨가시키지 않은 대조군과 첨가시킨 실험군으로 분류하였다. 동결방법은 난자를 1.5 M ethylene glycol (EG)에 2분 30초간 노출시킨후 5.5 M EG와 1 M sucrose가 첨가된 동결액에 20초간 노출시킨 후 Grid에 난자를 부착시킨후 직접 액체질소에 침지하여 동결하였다. 동결후 난자는 5단계로 융해를 실시한 후 정자와 체외수정을 시킨 후 수정된 난자는 modified P1 배약액에 124 h까지 발달율을 관찰하였고, 배양 후 발달된 배반포는 대조군과 실험군, 각각 4마리의 발정동기화된 recipient에 이식을 시행하였다. 배발달율은 대조군에 비해 실험군에서 4세포기 (48 vs. 84.4%), 8세포기 (34% vs. 70.6%), 상실배 (26% vs. 58.6%) 그리고 배반포 발달율은 (24% vs. 58.6%)로 높게 관찰되었다. 배아이식후 대조군과 실험군에서 각각 2 마리가 임신이되어 정상적인 산자를 분만하였다. 따라서 항동해제에 taxol의 첨가는 동결 융해후의 난자의 배발달율을 증진시킬 수 있었다..8%로 나타나 난할율 및 배반포 발생율에 있어서 융합조건에 따라 큰 차이는 없었으나 1.9㎸/cm, 30$\mu\textrm{s}$ 2회의 조건이 다른 조건들에 비하여 유의적으로 낮았다. 따라서, 체세포와 수핵란 세포질간의 융합율과 배반포 발생에 미치는 영향은 전압보다는 시간에 더 크게 받음을 알 수 있었으며, 이와 같은 결과에서 융합시 시간을 오래 주는 것보다 전압을 높이는 것이 수핵난자의 세포질에 상해를 줄이고 이후 배반포 발생에 유리할 것으로 사료되었다.면에서도 더욱 더 활발할 것으로 기대된다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며 큰 자기폭풍일수록 현저했다. 주상에서 관측된 이러한 특성은 서브스톰 확장기 활동이 자기폭풍의 발달과 밀접한 관계가 있음을 시사한다.se that were all low in two aspects, named "the Nonsignificant group". And the issues were high risk perception in general setting and

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.75-75
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• 2001

유리화 동결법은 동결중 ice crystal의 형성이 이루어지지 않으므로 난자의 동결보존을 위한 유용한 동결방법이다. 이전의 연구에서 마우스의 난자를 ethylene glycol과 electron microscope grid를 이용한 유리화 동결법으로 동결 융해한 결과 기존의 slow freezing 방법에서보다 높은 생존율과 배발달율이 나타남을 관찰하였다. 그러나 동물과 인간 난자를 이용한 연구를 통하여 난자의 경우 동결 융해후 염색체에 부착되어 있는 미세소관인 방추사가 온도의 변화에 매우 민감하여 염색체 이상성이 증가되는 것이 보고되었다. 이에 본 연구에서는 성숙난자를 유리화동결법에 의해 동결 융해후 난자의 염색체와 방추사의 이상성이 증가되는지 알아보고자 본 실험을 수행하였다. ICR mouse의 성숙란을 채취하여 연구목적에 따라 fresh한 대조군과 동결음해 시킨 실험군으로 분류하였다. 동결방법은 난자를 1.5 M ethylene glycol (EG)에 2분 30초간 노출 시킨후 5.5 M EG와 1M sucrose가 첨가된 동결액에 20초간 노출시킨 후 Grid에 난자를 부착시킨 후 직접 액체질소에 침지하여 동결하였다. 동결후 난자는 5단계로 융해를 실시한 후 생존된 난자는 tubulin 항체를 이용한 immunostaining 방법으로 방추사의 이상성을 관찰하였고, 염색체는 난자를 고정 후 10% Giemza로 염색 후 염색체의 수적인 이상성을 관찰하였다. 염색체 분석결과 염색체 이상 빈도는 대조군의 경우 19.6%, 동결융해군은 32.8%로 관찰되었다. 또 방추사의 이상빈도는 대조군의 경우 20.2%, 동결 융해군은 32.3%로 관찰되어 동결 융해후의 난자에서 염색체와 방추사의 이상 빈도가 증가됨이 관찰되었다.찰되었다. 배아이식후 대조군과 실험군에서 각각 2 마리가 임신이되어 정상적인 산자를 분만하였다. 따라서 항동해제에 taxol의 첨가는 동결 융해후의 난자의 배발달율을 증진시킬 수 있었다..8%로 나타나 난할율 및 배반포 발생율에 있어서 융합조건에 따라 큰 차이는 없었으나 1.9㎸/cm, 30$\mu\textrm{s}$ 2회의 조건이 다른 조건들에 비하여 유의적으로 낮았다. 따라서, 체세포와 수핵란 세포질간의 융합율과 배반포 발생에 미치는 영향은 전압보다는 시간에 더 크게 받음을 알 수 있었으며, 이와 같은 결과에서 융합시 시간을 오래 주는 것보다 전압을 높이는 것이 수핵난자의 세포질에 상해를 줄이고 이후 배반포 발생에 유리할 것으로 사료되었다.면에서도 더욱 더 활발할 것으로 기대된다. 배란후 72시간째에 초음파진단기를 이용하여 난소의 난포발달을 조사한 결과 , 대조구와 bFF처리구에 비해 AI처리구에서 발달난포가 유의적으로 많은 것을 확인하였다. 이상과 같은 결과로, Anti-inhibin serum은 한우 자체에서 분비하는 Inhibin을 특이하게 억제하여 Inhibin에 의해 억제되는 FSH분비가 촉진됨으로써 난포발달과 estrogen의 농도가 촉진되는 것으로 사료되어 anti-inhibin serum이 한우의 과배란유기 효과가 있는 것으로 사료된다.정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.째, 회복기 중에 일어나는 입자들의 유입은 자기폭풍의 지속시간을 연장시키는 경향을 보이며