• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.118-118
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• 2003

The aim of this study was to investigate the effect of glucose on embryonic development of mouse embryos. Two cell embryos were recovered from ICR female mice(3-4weeks) at 46~50 hrs after hCG 5 IU injection (mated just after hCG injection) and cultured in 50 $\mu m$ DMEM droplets supplemented with nothing (control: n=46), glucose 0.5mM (Group A; n=46) or glucose 3.15 mM(Group B; n=46) under mineral oil. All experimental media were supplemented with 20% human follicular fluid. Total blastocyst formation rates was lower (NS) in glucose groups (group A: 52.2% : B. 47.8%) than control group (60.9%). ZiB rates was the highest (P<0.05) in control (47.8%) than those in group A (21.7%) and B (28.3%). ZeB rates were the highest (NS) in group A (30.4%) than those in control (13.0%) and group B (19.6%). Blastocysts, cultured in group B (50.5), had the highest (NS) mean cell number compared with the others (control: 39.2 ; group A: (45.6). The ICM proportion (% ICM of total cells) in blastocysts cultured in group A (20.6%) was the highest (NS) than those of other tested groups (control: 15.2 ; group B: 13.9%). This study shows that a low dose of glucose added to culture medium increases the ICM proportion of blastocysts in mice.

• Journal of Embryo Transfer
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• v.17 no.1
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• pp.13-21
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• 2002

Intracytoplasmic Sperm Injection (ICSI) has been widely used fur both human infertility and basic research. However, the high incidence of chromosomal abnormality is severe problem in cattle. Various oocyte activation stimuli, therefore, were compared by assessment of developmental capacity and chromosome analysis. Motile sperm selected by Percoll-density gradient were treated with 5 mM dithiothreitol (DTT) and injected into an oocyte matured fur 24 h. Eggs were then allocated into 5 treatment groups. Group 1 (control), sperm injection was performed without any further activation stimuli to the oocytes. Group 2 (handled control), sham injection was performed without sperm. In Group 3, oocytes exposed to 5 (M ionomycin for 5 min at 39(C. Group 4. ionomycine + 1.9 mM demethylaminopurine (DMAP, 3 h) and Group 5, ionomycine + 3 h culture in Ml99 + DMAP. Cleavage and the later development rate in Groups 1, 2 and 3 were significantly (P<0.05) lower than those in Groups 4 and 5. The incidence of chromosomal abnormality in the embryos treated directly with DMAP after ionomycine was relatively higher than in the embryo of Group 3 h, delayed DMAP treatment. From this results DMAP caused to be arrested the release of the 2nd polar body, resulting in changes of chromosomal pattern. Therefore, the time interval between ionomycin and DMAP is a crucial role in bovine ICSI.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.33-33
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• 2003

This study was conduct to compare the efficacy to produce male and female somatic cloned piglets. Maturation of porcine COCs was accomplished by incubation in NCSU-23 medium supplemented with 0.6 mM cysteine, 10% porcine follicular fluid, 1mM dibutyryl cyclic adenosine monophosphate (dbc-AMP, Sigma, USA), and 0.1 IU/ml human menopausal gonadotrophin (hMG, Teikokuzoki, Japan) for 20h and then cultured without dbcAMP and hMG for another 18 to 24 h. Female and male fetal cells were isolated from each fetus, cultured in ES-DMEM medium containing 10% FCS. Enucleated oocytes were fused with fetal fibroblasts (passage 4 to 15). Reconstructed embryos were cultured in NCSU-23 with 4 mg/ml BSA under mineral oil at 39$^{\circ}C$ in 5% $CO_2$ in air. A total of 12,328 nuclear-transferred embryos (1- to 4-cell stage) were surgically transferred into 69 surrogate gilts. Three recipients aborted during the period of conception. Three gilts delivered eleven female piglets, and five recipients gave rise to birth 22 male piglets. The average birth weigh of the cloned piglets was 1.52 kg (1.38~1.83 kg) in female piglets and 0.84 kg (0.45~1.25 kg) in male piglets. Alive cloned pigs was seven in female piglets (63.6%) and four in male piglets (18.2%). The other two recipients is ongoing. This study suggests that female-derived fetal cell as a nuclear donor has more capability on production of cloned piglets than male.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.3
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• pp.94-100
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• 2015

Objective: The goal of this study was to ascertain optimal assisted hatching (AH) method in frozen embryo transfer. We compared the effect of depending on whether mechanical or laser-AH was performed before or after the vitrification of embryo development rate and blastocyst cell numbers. Methods: In order to induce superovulation, pregnant mare's serum gonadotropin followed by human chorionic gonadotropin were injected into 4- to 5-week-old female mice. 2-cell embryos were then collected by flushing out the oviducts. The Expanded blastocysts were recovered after the collected embryos were incubated for 48 hours, and were then subjected to artificial shrinkage (AS) and cross-mechanical AH (cMAH) or quarter-laser zona thinning-AH (qLZT-AH) were carried out using the expanded blastocysts before or after vitrification. After 48 hours of incubation, followed by vitrification and thawing (V-T), and blastocysts were fluorescence stained and observed. Results: The rate of formation of hatched blastocysts after 24 and 72 hours of incubation was significantly higher in the AS/qLZT-AH/V-T group than in the other groups (p<0.05). The cell number of the inner cell mass was higher in AS/V-T/non-AH and AS/V-T/cMAH groups than those of others (p<0.05). In the control group, the number of trophectoderm and the total cell number were higher than in the AS-AH group (p<0.05). Conclusion: The above results suggest that AS and AH in vitrification of expanded blastocysts lead to the more efficient formation of hatched blastocysts in mice.

• Journal of the Korean Institute of Oriental Medical Informatics
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• v.11 no.1
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• pp.52-57
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• 2005

Objectives : Recurrent pregnancy loss occurs in approximately 0.5-3% of women. There are many studies concerning immunological factor recently. Therefore, aim of this study is to examine the latest trend of researches concerning recurrent pregnancy loss, and controlled experiment on animals about antiphospholipid antibody. Method : We referred Pubmed site by using searching word of 'recurrent pregnancy loss' (Limits : 2000.1-2004.3, animal) Results and conclusions : 1. We searched 29 papers. Immunological factor : 18 ( about antiphospholipid antibody : 10 ), Chromosomal abnormality : 6, The others : 5 2. In five papers about controlled experiment on animals, (1) Materials : 8-12 weeks old mice / 11.5-day old-rat embryos / New Zealand rabbit (2) Inductions : inject intraperitoneally with human IgG containing antiphospholipid antibodies / culture in a solution of 1 ml medium which contained IgG purified from sera of women / inject intradermally with cardiolipin (3) Treatments : inject intraperitoneally with complement component before / culture in a solution of 1 ml medium which contained IgG purified from sera of women with SLE and RPL or from healthy women / inject intradermally with TFX, 0.9% NaCl (4) Measurements : weight fetuses and placentas, calculate frequency of fetal resorption / after culture, examine the embryos / examine platelet counts, APTT and numbers of live and dead newborns, resorbed fetuses, body mass, newborn viability and survival rates.

• 한국전산유체공학회:학술대회논문집
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• 2008.03b
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• pp.6-9
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• 2008

Analyzing the characteristics of blood flow in the blood vessels is very important to diagnose the circulatory diseases. In order to investigate the hemodynamic characteristics in vivo, the measurements of blood flows inside the extraembryonic arterial and venous blood vessels of chicken embryos were carried out using an in vivo micro-PIV technique. The circulatory diseases are closely related with the formation of abnormal hemodynamic shear stress regions, thereby it is important to get blood velocity and vessel's morphological information according to the vessel configuration and the flow conditions. In this study, the flow images of RBCs in blood vessels were obtained using a high-speed CMOS camera with a spatial resolution of approximately 14.6${\mu}$m${\times}$14.6${\mu}$m in the whole circulation network of blood vessels. The blood flows in the veins and arteries show steady laminar and unsteady pulsatile flow characteristics, respectively. The mean blood flows merged (in veins) and bifurcated (in arteries) smoothly into the main blood vessel and branches, respectively, without any flow separation or secondary flow which accompanying large variation of shear stress. Vorticity was high in the inner regions for both types of vessels, where the radius of curvature varied greatly. The instantaneous flows in the arterial blood vessels showed noticeable pulsatility due to the heart beat, and the main features of the velocity waveforms, including pulsatile shape, retrograde flow, mean velocity, maximum velocity and pulsatile frequency, were significantly dependent on the pulsatile condition which dominates the arterial blood flow. In near future, these in vivo experimental results of blood flow measured in various extraembryonic blood vessels would be very useful to understand the hemodynamic characteristics of human blood flows and various blood flow researches for clinically useful hemodynamic discoveries as well.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.3
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• pp.142-147
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• 2011

Objective: This study was performed to evaluate testicular sperm chromatin condensation using aniline blue-eosin (AB-E) staining and its effects on IVF-ET. Methods: Chromatin condensation was analyzed using AB-E staining in 27 cases of testicular sperm extraction. There were 19 cases of obstructive azoospermia (OA) and 8 cases of non-obstructive azoospermia (NOA) in IVF-ET. Mature sperm heads were stained red-pink whereas immature sperm heads were stained dark blue. The percentage of sperm chromatin condensation was calculated from the ratio of the number of red-pink sperm to the total number of sperm analyzed. Results: The overall percentages of chromatin condensation in OA and NOA were $31.1{\pm}11.2%$ and $26.3{\pm}14.4%$, respectively. The fertilization rate was significant higher in OA than NOA ($p$ <0.05); however, the rates of good embryos and clinical pregnancy did not show statistical differences. In OA and NOA, statistical differences were not observed in the rate of chromatin condensation, fertilization, good embryos, and clinical pregnancy between the pregnant group and non-pregnant group. Conclusion: Chromatin condensation is less stable than OA and showed a low fertilization rate in NOA. While there were no significant differences in chromatin condensation results between NOA and OA, we propose that a pattern of decreased chromatin condensation in NOA is one of the factors of low fertilization results requiring further study.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.2
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• pp.258-264
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• 2000

Production systems for cloned pigs are very important not only for an increase in production of superior animals but also for the production of knockout animals with organs that do not contain antigens for xenotransplantation or to analyze functions of isolated human genes. At present, however, effective systems have not been developed. We have tried to produce cloned pigs by transfering cultured cells into enucleated oocytes and obtained some cloned embryos. To develop a production system for cloned pigs, the basic technologies needed to support such an effort must be improved.

• 대한생식의학회:학술대회논문집
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• 2006.11a
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• pp.121.2-122
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• 2006

• 대한생식의학회:학술대회논문집
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• 2002.11a
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• pp.97-97
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• 2002