• Journal of Embryo Transfer
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• v.18 no.3
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• pp.249-255
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• 2003

These studies were performed to improve the reproductive efficiency of gilts and we investigated the effects of puberty periods, first mating time and backfat thickness and will adapt to these results for early selection of excellent gilts. The main results were as follows; 1. First heats on birth season were showed 194.14 day, 163.25 day, 160.25 day and 157.92 day at birth of spring, summer, autumn and winter, respectively and birth of spring was significantly latest among other seasons (p<0.01). 2. First service on birth season were revealed 222.05 day in spring, 193.00 day in summer, 199.20 day in autumn and 190.11 day in winter. birth of spring was significantly latest among others (p<0.01). 3. First heat period of cadidated gilt had 13∼16 mm backfat thickness was 180.32 day, 171.24 day in 17∼20 mm and 162.20 day in 21∼23 mm and was showed delay in thin backfat gilts. There was no differences among backfat thickness. 4. First service of cadidate gilt had 13∼16mm backfat thickness was 211.12 day, 202.43 day in 17∼20 mm and 195.43 day in 21∼23 mm and was showed delay in thin backfat gilts. There was no differences among backfat thickness. 5. The litter size were 9.64 in gilts under 160 day of first heat, 10.14 in 161∼180 day, 9.56 in 181∼200 day and 9.13 in over 201 day. There showed the largest litter size in 161∼180 day of first heat but was no differences. 6. The litter size in gilts under 180 day of first service was 9.13, 9.75 in 181∼200 day, 10.13 in 201∼220 day and 9.45 in over 221 day. There showed the largest litter size in 201∼220 day of first service but was no differences. 7. The litter size of gilt had 13∼16 mm backfat thickness on first service was 9.33, 9.81 in 17∼20 mm and 10.17 in 21∼23 mm and was showed delay in thin backfat gilts. There was no differences among backfat thickness.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.69-73
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• 2003

This study was undertaken to investigate the effect of three different porcine IVM media, TCM-199, mSOF and NCSU-23 on early development of porcine spontaneous parthenogenotes. Spontaneous parthenogenotes were induced by a prolonged culture of porcine oocytes in each medium. In Experiment 1, oocytes derived from gilts were matured in three IVM media and maturation of oocytes was evaluated by the status of chromatin configuration. Oocytes matured in mSOF, NCSU-23 and TCM-199 showed no significant difference (P>0.05) in maturation. Maturation rates at 48h after IVM were 83.1$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (TCM-199). In Experiment 2, pronucleus formation and development to 6~8 cell stage of pig oocytes activated spontaneously. Pronucleus formation, cleavage rate and development to 6~8 cell embryos of porcine spontaneous parthenogenotes were assessed at 55~58 h, 96 h and 168h after IVM, respectively. Pronucleus formation (5.4$\pm$2% and 3.7 $\pm$ 1% vs 1.7 $\pm$ 3%) and development to the 6$\pm$8 cell (3.2$\pm$3% and 4.0$\pm$1% vs 1.4$\pm$3%) was significantly (P<0.05) higher in mSOF or NCSU-23 than TCM-199. In conclusion, the present study showed that oocytes matured in mSOF and NCSU-23 were spontaneously activated with higher frequency.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.1-14
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• 2003

This study was conducted to investigate that the immature and mature oocytes of porcine can be cryopreserved by vitrification. Oocytes were centrifuged to polarize the cytoplasmic lipid droplets. The lipids were removed from cytoplasm by micromanipulation. Delipated oocytes were centrifuged after being preincubated with cytochalasin B(CB) fer 10 min, and lipid droplets were removed. Centrifuged oocytes were treated with CB and centrifuged to polorize lipid droplets but not delipated and control oocytes is not-treatment. Oocytes of three types were vitrified in electron microscope(EM) grids. The results of survival, maturation and cleavage rates were as follows. 1 The survival rates of immature oocytes were 15.1%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated wassignificantly higher than Centrifuged and Control(P<.01). 2. The survival rates of mature oocytes were 12.21%, 0% and 0% in the Delipated, Centrifuged and Control after vitrification, respectively, and its rate of Delipated was significantly higher than Centrifuged and Control(P<.01). 3 The maturation rates of immature oocytes were 37.5% and 68.9% for metaphase II in the Delipated after vitrification and Non-vitrification, respectively, and its rate of Non-vitrification was significantly higher than Delipated after vitrification(P<.01). 4. The cleavage rates of immature oocytes were 12.5%, 0%, 0% and 56.1% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05). 5 The cleavage rates of mature oocytes were 25.0%, 0%, 0% and 67.9% in the Delipated, Centrifuged, Control after vitrification and Non-vitrification, respectively. It's rate of Delipated was higher than Centrifuged and Control, but there were no significant difference, and its rate of Non-vitrification was significantly higher than Delipated, Centrifuged and Control(P<.05).

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.45-50
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• 2015

The purpose of this study is to produce wanted sex progeny of genetically confined White Hanwoo (albinism) with preselected sex sperm. One bull of White Hanwoo was chosen for semen donor and X sperm was sorted by MoFlo XDP cell sorter. To compare the pregnancy and birth rates, KPN straw was used as control, total number of unsorted sperm was $20{\times}10^6/straw$. Sexed X frozen semen with $20{\times}10^6$ cells or $4{\times}10^6$ cells per straw were in seminated twice on Hanwoo heifers. The abnormality of the sexed X semen was $24.9{\pm}7.31%$ and distal reflex abnormality of mid piece was significantly (p<0.05) higher (11.7%) compared with that of KPN 768 (5.6%). There were no differences on the pregnancy and birth rates between $2{\times}10^6$ cells or $4{\times}10^6$ cells of X-sperm but KPN semen showed significant differences (p<0.05). The pregnancy rates of KPN 768, $2{\times}10^6$ cells and $4{\times}10^6$ cells X-sperm of White Hanwoo cattle were 85.0%, 26.3% and 50%. The birth rates were 80.0%, 15.8% and 21.4%, respectively. The female offspring rates of KPN 768, $2{\times}10^6$ cells and $4{\times}10^6$ cells X-sperm of White Hanwoo cattle were 43.8%, 100% and 100% (p<0.05). These results indicated that sex sorted White Hanwoo could be used for the production of wanted progeny with $2{\times}10^6$ cells/straw for AI. To increase the efficiency of calf production, the sperm number of sex sorted semen will be optimized for sex selection of White Hanwoo progeny.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.183-188
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• 2015

This study analyzed the coat color-related genes of MC1R, ASIP, ECA3-inversion, and STX17 of 1,462 Jeju horses administered by the Jeju Special Self-Governing Province. This was done to investigate the distributional characteristics of coat color-related genes in the Jeju horse group and the changes of its coat color-related genes by generation. The genotype frequency of the MC1R gene of $E^+/E^+$ and $E^+/E^e$ related to black coat color was 0.122 and 0.447, respectively, while $E^e/E^e$ of the chestnut genotype was 0.429. The genotype frequency of the ASIP gene of $A^A/A^A$, $A^A/A^a$, and $A^a/A^a$ was 0.46, 0.448, and 0.091, respectively, where the genotype frequency of $A^a/A^a$ turned out to be relatively low. The To/To and +/To genotype that manifests the Tobiano shape was 0.001 and 0.119, respectively, with the share of Tobiano shape around 12%. The genotype frequency of G/G and G/g of STX17 related to grey coat color was 0.002 and 0.680, respectively, with the share of grey horses among the Jeju horse group at 68.2%. As for the change of coat color genes by generation, no large changes were observed in the MC1R and ASIP genes. In ECA3-inversion, the To allele that manifests Tobiano significantly decreased following the generational change (p<0.05), while the STX17 G allele related to grey coat color significantly increased following the generational change (p<0.05). It will be necessary to examine the coat color genes when selecting breeding horses so that the diversity of coat colors among the Jeju horse group can be maintained.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.175-181
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• 2015

The objective of this study was to monitor health conditions of genetically identical somatic cells cloned Korean white cattle, endangered indigenous cattle (EIC) and indigenous cattle (IC) by analysis of hematologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and one live cattle were delivered by caesarean section. The cloned Korean white cattle were genetically identical to the nuclear donor cattle. As a result, the mean values of RBC and platelet of cloned cattle and white cattle were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between cloned cattle and IC of the same age (1~2 years) showed the statistical significance (P<0.05). Also, in the WBC of Korean white cattle, the estimated values were decreased according to the age from $12.0{\times}10^3/{\mu}l$ under 1 year to $11.0{\times}10^3/{\mu}l$ over 1 years respectively. Although clone-cattle had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned cattle were not different when compared to reference range. This study suggests that cloned Korean white cattle derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological parameters. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.7-15
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• 2015

The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilization ability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezing solution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawed such as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double staining for mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDA staining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in 5 and 10 mM ($61.1{\pm}1.5%$,$64.7{\pm}2.0%$) of nicotinic acid than other groups (0 mM, $52.1{\pm}2.3%$; 20 mM, $47.8{\pm}5.1%$, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid ($26.1{\pm}1.8%$, $24.9{\pm}1.5%$) than other groups (0 mM, $35.3{\pm}0.8%$; 20 mM, $36.5{\pm}1.9%$, P<0.05). The live sperm with mitochondrial integrity was significantly higher in 5 and 10 mM ($84.2{\pm}3.6%$, $88.4{\pm}2.3%$) of nicotinic acid than other groups (0 mM, $77.3{\pm}4.4%$; 20 mM, $73.3{\pm}3.6%$, P<0.05). Blastocyst rate of in vitro development was significantly higher in 10 mM ($17.0{\pm}1.3%$) of nicotinic acid than other groups (0 mM, $9.4{\pm}0.5%$; 5mM, $12.6{\pm}0.8%$; 20 mM, $5.0{\pm}1.0%$, P<0.05). Moreover, total cell number was higher in 5 and 10 mM ($53.6{\pm}2.9%$, $57.9{\pm}2.8%$) of nicotinic acid than other groups (0 mM, $41.0{\pm}1.4%$; 20 mM, $23.2{\pm}2.8%$, P<0.05). Hydrogen peroxide in embryos was lower in 5 mM nicotinic acid ($0.7{\pm}0.1%$) than other groups (0 mM, $1.0{\pm}0.1%$; 10mM, $0.9{\pm}0.0%$; 20 mM, $1.4{\pm}1.0%$, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and quality of spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.265-271
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• 2014

Implementation of smart embryo technologies in cattle e.g. ovum pick-up followed by in vitro embryo production (OPU-IVP). Seasonal variation is important factor for follicular growth, oocytes quality, quantity and developmental competence. Therefore the aim of present study was carried out to investigated whether the seasons (hot and cool) effect on follicular development, oocyte recovery and subsequent embryo development. Follicular oocytes were aspirated from Korean native cows (Hanwoo) by the ovum pick-up (OPU) method, which was performed 24 times during two different seasons, the hot (July to September) and cool (October to December), from OPU donors. The recovered oocytes were classified according to morphological categories and used for in vitro embryo production (IVEP). The mean number of total follicles was significantly higher (p<0.05) during the hot season ($18.32{\pm}2.26$) compared to cool season ($15.41{\pm}3.34$). Furthermore, seasons did not significantly effect on the number of oocytes recovered (hot season: 41.16% vs. cool season: 46.14%). However, the average number of Grade A oocytes was significantly greater during hot ($1.75{\pm}1.86$) season compared to the cool season ($1.00{\pm}1.46$), but there was no significant difference of other grades oocytes. The cleavage rate (hot: 66.67% vs. cool: 63.3%) and embryo development (hot: 58.95% vs. cool: 56.97%) did not differ significantly between the seasons. In conclusion, the results of present study suggest that the season (hot and cool) does not have effects on the oocyte recovery and embryo developmental competence of in vitro cultured embryos.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.241-248
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• 2014

This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by PMSG and hCG. Two-step EG, DMSO and 4-step EG, DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in EG and DMSO was significantly higher than 4~8 cell (65.4% versus 61.2%, 81.1% versus 72.5%) (p<0.01, p<0.01), but the development rates of 4~8 cell embryos in EG and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4~8 cell embryos in EG were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in EG(77.0% versus 64.4%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The development rate from morular to hatching blastocyst, however, was sinificantly higher in EG than in DMSO during 48 hr (p<0.01). The survival rate of 4~8 cell embryo was 62.5% in EG and 73.3% in DMSO. The development rates of embryo in EG were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. The survival rate of embryo in 2 cell stage was higher than in 4~8 cell stage, and EG appears more effective cryoprotectant than DMSO because EG showed better development rates of embryos in 2 and 4~8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.313-319
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• 2014

Investigations for hematologic values and the differential count of WBC for Korean indigenous cattle (KIC) and Stripped Cattle (SC) are rarely performed. Therefore, when the index of complete blood counts (CBC) analysis of KIC and SC were requested, it had many difficulties to make the results for blood condition since the standard hematologic values of KIC and SC are lacking. The objective of this study was to investigated the hematologic values and the differential count of WBC for blood of total 19 striped cattle (SC) and 187 KIC as a control by estimation analysis of hematologic characteristics. As a result, the mean values of RBC and platelet of KIC were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between KIC and SC of the same age (2~3 years) showed the statistical significance (P<0.05). Also, in the WBC of KIC, the mean values were decreased according to the age from $13.8{\times}10^3/{\mu}l$ under 1 year to $9.5{\times}10^3/{\mu}l$ over 5 years. In the differential count of WBC of KIC, it showed generally the rates of 46.2% lymphocyte and 36% segmented neutrophil. Additionally, in comparative analysis between pregnant and non-pregnant group of KIC, the mean values of Hg and HCT in pregnant group were significantly decreased(P<0.05). In conclusion, data obtained from this study may be valuable as a standard for interpretation of the results in hematologic analysis of KIC. Result of this study will be used for establishing reference range for hematologic analysis in SC.