• Journal of The Korean Society of Grassland and Forage Science
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• v.19 no.4
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• pp.297-302
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• 1999

Cotyledonary abnormalities were observed in secondary somatic embryos which were developed from calli cultured on MS medium with various concentrations of 2,4-D. In MS medium containing hormone-free or $0.1mg/{\ell}$ 2,4-D, the frequency of normal embryo formation with two cotyledons were above 57%. According to concentration of 2,4-D increment the frequency of normal embryos were decreased. In MS medium containing $4mg/{\ell}$ 2,4-D, the frequency of normal embryo formation was just 10%. The rate of germination was as follows; 37.7% of somatic embryos had one cotyledon, 85% two cotyledons, 38% three cotyledons, 35% four cotyledons, 25% five cotyledons and 29% trumpet-like cotyledons. About 80% of the embryos with two cotyledons were converted into normal plants, but one, three or four cotyledons were only 6.8~10%. The five or trumpet-like embryos were not developed into normal plants.

• Journal of Embryo Transfer
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• v.11 no.3
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• pp.225-231
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• 1996

This study was conducted to investigate the effects of preincubation time, concentration and exposure time of sperm on in vitro fertilization of porcine follicular oocytes rnatured in in vitro. The results obtained are as follows ; 1. Effect of preincuhation time for porcine sperm capacitation on in vitro fertilization in medium with heparin was investigated. Normal fertilization rate was highest in 15 min(26.4%). However, there were no significant differences among preincuhation times of 5~90 min, 2. Normal fertilization rates of sperm concentrations were 17.0~26.5%, and normal fertilization rate from l$\times$ l05cell /ml concentration was also higher than those of other sperm concentration. 3. Normal fertilization rates of sperm exposure time of 4, 8, 12, 16 and 20 hours were 6.1, 20.8, 27.8, 25.0 and 26.7%, respectively. Normal fertilization rate from sperm exposure time of 12 hours was also higher than that of other sperm exposure times.

• Horticultural Science & Technology
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• v.31 no.4
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• pp.483-489
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• 2013

The abnormal seedlings, a common physiological anomalies, emerged during embryo rescue severely restricted grape breeding. To enhance the efficiency of the seedless grape breeding by reducing the production of abnormal seedlings in the course of embryo rescue, we investigated the effects of genotype, media type, embryo style, pre-chilling on the deformity rate of the abnormal seedlings during embryo rescue. The abnormal seedlings were firstly classified into seven categories based on their morphology. Our results indicated that the emergence of abnormal seedlings was highly dependent on the female parent genotype. Polyembryony was advantageous to diminish the number of abnormal plantlets and the germination rate of embryo was 100%. We also found that pre-chilling treatment could reduce the number of abnormal plantlets and promote the embryo germination. The abnormal plantlets were reduced significantly by the addition of $ZnSO_4$ $10{\mu}mol{\cdot}L^{-1}$ or mashed-banana $500mg{\cdot}L^{-1}$ to either embryo development or germination media. Transferring the abnormal seedlings onto the suitable fresh media in 4 weeks after embryo germination provided an effective way to transform them into normal seedlings.

• Journal of Embryo Transfer
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• v.5 no.1
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• pp.21-27
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• 1990

The technique for maturation of follicular oocyte has been devised to provide such a low cost and in ptentifut number supply of bovine embryo. Some of problems concerning production of bovine embtyo in vitro were discussed in this paper. Bovine follicular oocytes cultured in vitro achieved normal fertilization but cleavage rates to blastocyst were low compared to the oocyte matured in vivo. It has been concluded that a deficient cytoplasmic maturation occurs in the oocytes matured in vitro. These results indicate that the studies for maturation of bovine follicular oocytes in vitro need improvement of culture conditions and to define the characteristics that might be indicative of healthy oocyte.

• Clinical and Experimental Reproductive Medicine
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• v.51 no.1
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• pp.63-68
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• 2024

Objective: This study was conducted to investigate the impact of previous delivery mode on pregnancy outcomes in patients with secondary infertility after frozen-thawed embryo transfer. Methods: This prospective observational study included 140 patients experiencing secondary infertility. Of these, 70 patients had a previous cesarean delivery (CD), while the remaining 70 patients had a previous normal vaginal delivery (NVD). The primary outcome was the implantation rate. The secondary outcomes included rates of clinical pregnancy, chemical pregnancy, miscarriage, and ectopic pregnancy. Results: The comparison of all fertility outcomes between the two groups revealed no statistically significant differences. The implantation rate was 40.4% in the CD group and 41.7% in the NVD group (p=0.842). The clinical pregnancy rate was 50% in the CD group and 49.3% in the NVD group (p=0.932), while the chemical pregnancy rate was 14.6% in the CD group and 19% in the NVD group (p=0.591). The miscarriage rates in the CD and NVD groups were 20% and 17.6%, respectively (p=0.803). One case of tubal ectopic pregnancy occurred in the NVD group (1.4%). Conclusion: The mode of prior delivery did not significantly impact pregnancy outcomes following frozen-thawed embryo transfer.

• Journal of Embryo Transfer
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• v.4 no.1
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• pp.41-45
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• 1989

Total thirty of flushing were attempted on day 4 to 15 of estrus cycle with S heifers and 9 cows by nonsurgical method. The flushed or recovered rate among flushings was 86.7% (26/30) or 88.5% (23/26), respectively. There was no difference in the recovered rate between heifers (85.7%,6/7) and cows (89.5%, 17119). The embryo was recovered on day 4 to 15 of estrus cycle from the donors in natural heat without any technical difficulties.The I2FG Foley catheter used for pubertal heifers had sometimes plug in it with uterine mucus during flushing of uterine horn. But the problem could be overcomed by pumping the catherter with fluthing solution or by changing the catheter. Three normal embryos were recovered from 3 pubertal (10-11 month old) heifers. The rate of normal and abnormal eggs was 60.9% (14123) and 39.1% (9/23), respectively. The abnormal eggs were on degenerating except one unfertilized egg and were mostly recovered from heifers or cows flushed consecutively during the estrus cycle. The developmental states of normal embryos were l6-cells on day 5, 32-cells on day 6, compacted-morula on day 7, early-to expanded-blastocyst on day 8-to 9, and hatching-to hatched-blastocyst on day 10 to 11 of estrus cycle. The stage of embryos on day 8 to 10 showed varities among donors. On day 8 to 9 of estrus cycle hatching-blas tocyst was recovered from some donors.

• Journal of Acupuncture Research
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• v.35 no.4
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• pp.200-206
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• 2018

Background: To evaluate whether ${\geq}3$ adjunct acupuncture sessions accompanying embryo transfer, increases the chance of pregnancy amongst post-myomectomy women aged ${\geq}35$ years. Methods: This was a prospective study carried out at Nordica Fertility Center. Following written informed consent, 75 patients undergoing assisted reproduction therapy and who had good quality embryos, were age-matched and grouped into post-myomectomy (n = 24) and normal women who had no evidence of fibroids or previous myomectomy (n = 51). Between 1 and 3 sessions of acupuncture were performed on 6 post-myomectomy and 19 infertile women who had not undergone myomectomy, while > 3 acupuncture sessions were performed on 18 post-myomectomy and on 32 normal patients, approximately 25 minutes before and after embryo transfer. Results: A positive pregnancy test was defined as ultrasonographic evidence indicating presence of a fetal sac 6 weeks after embryo transfer. Of the 5 post-myomectomy women who were pregnant, only 1 (20.0%) received 1-3 adjunct acupuncture sessions whilst the remaining 4 (80.0%) received > 3 acupuncture sessions. Of the 11 normal pregnant women, 5 (45.4%) received 1-3 adjunct acupuncture sessions while 6 (54.5%) received > 3 adjunct acupuncture sessions. Conclusion: Pregnancy rates in infertile post-myomectomy women may be improved by > 3 adjunct acupuncture sessions.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.229-234
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• 1994

This experiment was carried out to develop the model system for mass production of biomedical and nutritional proteins (human proteins) through mamraary gland of the transgenic cattle produced by gene manipulation and embryological technologies. Human growth hormone gene fused with rat $\beta$-casein gene promoter was microinjected into pronuclei of one cell bovine embryos produced by in vitro fertilization. After microinjection, embryos were cultured in vitro for 6 or 7 days. Twenty embryos reaching to blastocysts were transferred to 10 beef recipients, each receiving two embryos. Recipients were diagnosed for pregnancy by rectal palpation at 76 days after embryo transfer. One of them was pregnant to term and produced a female calf weighing 21 kg at 280 days following embryo transfer. DNA was extracted from umbilical cord tissue and blood of calf born for confirming gene insertion. As determined by Southern hybridization, the transgene was not found.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.185-191
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• 2013

The purpose of this study is to improve production efficiency of vitrified-thawed transgenic bovine embryos. Transgenic bovine embryos were produced by injection of FIV-GFP lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization. EGFP-expressing transgenic bovine blastocysts were cultured in serum-containing and serum-free medium. These blsatocysts were vitrified by pull and cut (PNC) container made with 0.25 cm plastic straw. Results indicate that total developmental rates of normal IVF embryo cultured in serum-containing and-free medium into blastocyst were not significantly different (22.3 vs 21.5%) and those of GFP-expressing transgenic bovine embryo into blastocyst showed no significant difference between serum-containing (13.9%) and-free medium (13.1%). However, developmental rate of GFP transgenic embryo was significantly (P<0.05) lower than its of normal IVF embryos. In additional study, we vitrified GFP transgenic normal bovine blastocysts using PNC vitrification method. Survival rate of vitrified-thawed GFP transgenic blastocyst (23.1%) was significantly (P<0.05) lower than its of normal blastocysts (68.9%). Although, survival rate of vitrified-thawed GFP transgenic blastocyst was lower than its of normal blastocyst, our result may suggested that PNC vitrification method is feasible to cryopreserve transgenic embryos. Our next plan will be the production of GFP express transgenic bovine derived from vitrified-thawed embryos using PNC method.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.3
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• pp.167-178
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• 2002

Objective : The present study was performed to investigate whether apoptosis occur in human embryos by annexin staining and detect the expression of Fas, Fas-ligand (FasL), Bax, and Bcl-2 in human fragmented embryos derived from IVF-ET by immunofluorescence and Western blot analysis. Materials and Methods: Using annexin staining, immunofluorescence and Western blot analysis on normal and fragmented embryos, we were able to detect apoptotsis and apoptotic gene products in fragmented embryos. Result: Phosphatidylserine (PS) translocation, the marker for apoptosis, were detected frequently in fragmented embryos. Bcl-2 and Bax protein were detected in both fragmented and non-fragmented embryos. When fragmented embryos compared to normal embryos, immunofluorescent intensity of Bcl-2 tended to be lower in fragmented embryos. Bax gene expression increased in the fragmented embryos compared to the normal embryos. This result supports a model in which the molar ratio of Bcl-2 to Bax determines whether apoptosis induced or inhibited in human embryo. Fas was highly expressed in human preimplantation embryos but not FasL. It suggests that embryo may undergo apoptosis by binding with FasL produced by follicular or immune cells. Conclusion: The over expression of Bax and Fas will trigger apoptosis to lead embryo fragmentation and change embryo to be nonviable.