• 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.13 no.1
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• pp.11-19
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• 1986

Follicular flxid (FF) and their matched oocytes were obtained from 58 follicles of 27 women who underwent an in vitro fertilization (IVF) procedure with ovarian hyperstimulation by clomiphene citrate(n=8), hMG(n=9),FSH/hMG(n=10). Follicular aspiration was performed 36 hours after human chorionic gonadotropin administration. The concentcation of androstendione (ADD), testosterone (T) was correlated with hyperstimulation regimens, the morphology of the oocyte-corona-cumulus complex (OCCC), oocyte fertilization, and the incidence of pregnancy after embryo transfer. The results were as follows. 1. According to hyperstimulation regimens, there was no significant differance in FF ADD and T concentrations of the similar morphology of OCCC. 2. In clomiphene-treated and FSH/hMG-treated cycles, FF ADD concentrations of preovulatory oocytes were 43.09${\pm}$9.53 ng/ml and 59.46${\pm}$9.09 ng/ml, those of immature occytes were 96.98${\pm}$16.55 ng/ml and 116.13${\pm}$36.81 ng/ml, those of atretic oocytes were 246.5 ${\pm}$9.25 ng/ml and 634.25${\pm}$9.25 ng/ml respectively, reflecting the significant relationship between FF ADD level and morphologic maturity of OCCC (p<0.05). But in hMG-treated cycles, such relationship was not found (p>0.1). In clomiphene-treated and FSH/hMG-treated cycles, FF T concentrations of preovulatory oocytes were 11.37${\pm}$2.38 ng/ml and 11.68${\pm}$1.73 ng/ml respectively which were significantly lower than those of atretic oocytes (25.1${\pm}$7.50 ng/ml and 23.25${\pm}$0.95 ng/ml respectively) (p<0.05). But in all cycles, FF T concentrations of immature oocytes were not significantly different from those of preovulatory oocytes, artetic oocytes (p>0.1). 3. In hMG-treated and FSH/hMG-treated cycles, FF ADD concentrations of fertilized oocytes were 32.43${\pm}$4.09 ng/ml and 42.61${\pm}$4.82 ng/ml respectively which were significantly lower than those of non-fertilized oocytes (72.18${\pm}$17.31 ng/ml and 108.09${\pm}$17.32 ng/ml respectively) (p<0.05), but in clomiphene-treated cycles there was no significant difference (p>0.1). In FSH/hMG-treated cycles, FF T concentration of fertilized oocytes was 7.33${\pm}$1.06 ng/ml which was significantly lower than that of non-fertilized oocytes (20.3${\pm}$6.21 ng/ml) (p>0.02), but in clomiphne-treated and hMG-treated cycles there was no significant difference (p>0.1). 4. In all cycles FF ADD and T concentrations did not correlated with the success of pregnancy after embryo transfer. Above results suggested that FF ADD and T may play an important role in oocyte maturation and fertilization, but their relationship with the success of psegnancy was not found.

• The Journal of Korean Obstetrics and Gynecology
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• v.26 no.2
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• pp.120-137
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• 2013

Purpose: This study was to investigate status survey and cost of infertility patients in ${\bigcirc}{\bigcirc}$ Korean Medicine Hospital. Methods: From January 2012 to June 2012, 171 new patients were admitted to ${\bigcirc}{\bigcirc}$ Korean Medicine Hospital for treatment of infertility. We reviewed their medical records retrospectively and had telephone consultations to find out whether they were pregnant or not. Results: The mean age of outpatients was $32.72{\pm}4.08$ years and mean BMI of outpatients was $20.26{\pm}2.68$. 55.0% of patients who did not have childbirth or miscarriage. 82.5% of patients had normal menstrual period. 93.5% of patients visited OB/GYN, 33.3% were treated with ovulation induction, 18.7% underwent intrauterine insemination, and 18.1% underwent in vitro fertilization and embryo transfer. 99.7% were treated with acupuncture and moxibustion, 100% had taken Herbal Medicine. The mean treatment duration of outpatients was $8.84{\pm}8.17$ weeks, and the mean medical expense was $761,994{\pm}586,502$ won. It was found that 32.4% of patients were pregnant after treatment. Conclusions: We investigated status survey and cost of infertility patients. Further study about Korean medical treatments on infertility is required.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.57-64
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• 2003

Objective : We investigate the effects of multiple attempts of embryo transfer because of retained embryos in the catheter and of contaminated mucus on the transferred catheter. Materials and Methods: We respectively analysed data between November 1998 and August 2002 from 305 patients of 369 cycles who underwent IVF-ET. Of these patients, 47 patients of 50 cycles (Group 2) were required multiple trial of embryo transfer. They were compared with an age-matched control groups (Group 1) with female factor infertility. Pearson's $?^2$ and Fisher's tests were used to compare proportions between discrete variables. Noncategorical data were compared using t-test. Statistical significance was set at p<0.05. Results: Embryos were significantly more likely to be retained when catheter was contaminated with mucus (Group 1: 22.4%; Group 2: 44.0%). The clinical pregnancy rates, however, for the contaminated mucus or not, were 46.8%, 43.5% respectively. There was no significant difference clinical pregnancy rate between those who had all their embryos transferred at the first attempt (45.4%) and those who required more than one attempt (48.0%). Conclusions: Contaminated mucus in the catheter is associated with failed embryo transferred at the first attempt. Embryo transfers, however, that are repeated attempts do not adversely affect pregnancy rates following IVF-ET.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.4
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• pp.273-277
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• 2004

Intramural pregnancy is an unusual ectopic gestation located within the uterine wall, completely surrounded by myometrium and separate from the uterine cavity, fallopian tube, or round ligament. It is known to be difficult to diagnose, and associated with a high rate of uterine rupture. We report a case of intramural pregnancy in which early diagnosis was made and successful treatment was done by dilatation and curettage. Diagnostic laparoscopy confirmed the absence of uterine rupture during the procedure. Therefore, conservation of fertility can be possible with early diagnosis of intramural pregnancy. To our knowledge, this is the first case report of intramural pregnancy following IVF-ET in Korea.

• Korean Journal of Agricultural Science
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• v.42 no.4
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• pp.355-359
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• 2015

The cause of infertility is either fertilization failure or early embryonic death. The aetiology may involve a combination of many factors, e.g. genetic factors, abnormalities in the gametes nutritional disorders, inadequate luteal function, and delayed ovulation. This study was conducted to investigate the effect of microorganisms collected from uterus of Hanwoo cattle on early embryonic development. Microorganisms isolated from the uterus of Hanwoo cattle included Bacillus cereus (Bc), Bacillus thuringiensis (Bt), Staphylococcus warneri (Sw) and Enterococcus faecalis (Ef). When cultured with Bc, Bt, Sw, and Ef, oocytes were not developed into a blastocyst in vitro. The proportion of blastocyst was dramatically increased after reducing the number of microorganisms ($10^4CFU/ml$). Interestingly, the proportion of blastocyst was decreased by adding the Sw and Ef. These results indicate that among intrauterine microorganisms, Sw and Ef strains negatively affect theearly embryonic development, thereby aggravate the rates of implantation and pregnancy. These findings will provide useful information for association studies in other pig populations.

• Clinical and Experimental Pediatrics
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• v.46 no.3
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• pp.224-229
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• 2003

Purpose : To examine various neonatal outcomes and perinatal factors resulting from assisted reproduction compared to that of spontaneous conception. Methods : This is a retrospective study. The control cases were all twins of spontaneous conception born between periods from January 1995 to June 2000. The study cases were identified from twins conceived by assisted reproduction in the same time peried. A total of 460 sets of twins consisted of 250 twins of spontaneous conception and 156 twins of assisted reproduction were studied. The primary outcomes were neonatal morbidity and mortality and the secondary outcomes were perinatal factors including number, length and cost of hospitalization for the delivery. Results : No differences were seen in various neonatal factors including gestational age, birth weight and incidences of respiratory distress syndrome, patent ductus arteriosus, necrotizing enterocolitis, hyperbilirubinemia, sepsis, intraventricular hemorrhage and the length of hospitalizations. Lower one minute and five minute Apgar scores and frequently encountered electrolyte abnormalities were observed in neonates of assisted reproduction. In general, the second twin of assisted reproduction had increased incidences of respiratory distress syndrome, sepsis and necrotizing enterocolitis than the first twin. Increased frequencies of preterm labor, hospitalization and elective cesarean section were seen among mothers who underwent artifical conception. However, overall hospital costs in terms of mothers hospitalization for the delivery and neonates hospitalization did not show differences. Conclusion : Assisted reproduction twins had similar neonatal morbidities, mortalities and perinatal morbidities compared to those born by spontaneous conception.

• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.27-34
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• 1996

The objective of this study was to examine the cell number of Total, ICM and TE cells of bovine blastocysts according to development progression cultured in CR1 medium, which was reported as successfully supporting medium for preimplantaion bovine embryo development to the blastocyst stage, by differential labelling of the nuclei with immunosurgery and polynucleot-ide-specific fluorochromes. Blastocysts were obtained at day 8 after in vitro fertilization and classified to early, middle, expanded stage according to the developmental morphology; blastocoel expansion and zona thickness. Also, bias tocysts in the same category were divided into two parts to check the Total cell number by using bisbenzimide only and ICM, TE and Total cell number by using immunosurgery and two polynucleotide-specific fluorochromes. 1) The development rate of blastocysts at day 8 after in vitro fertilization was 29.3% and classified bIas tocysts to early, middle, expanded and hatching stage were 8.7, 9.9, 7.6 and 3.1%, respectively. 2) The numbers of total blastomere using bisbenzimide in the classified blastocysts to early, middie and expanded were 46.9${\pm}$8.6, 66.2${\pm}$12.5 and 122.8 ${\pm}$ 14.4, respectively. This indicated that CR1 is a appropriate culture medium for bovine embryo development. 3) The count of ICM and TE cell number by using differential labelling with immunosurgery and polynucleotide-specific fluorochromes in the classified blastocysts to early, middle and expanded; ICM cell numbers of were 12.8${\pm}$5.9, 26.3${\pm}$8.4 and 35.5${\pm}$15.0, respectively and TE cell numbers were 30.5${\pm}$5.0, 4 41.3${\pm}$8.2 and 81.1${\pm}$13.4, respectively. These results presented that the increase of ICM and TE cell numbers averaged two and three doublings between early and expanded blastocyst stage and also total cell number counted from ICM nuclei and TE nuclei by using differential label-ling showed the increase pattern with development advance level and the results were similar to total cell number obtained from bisbenzimide treatment only. Therefore, the differential labelling of ICM and TE nuclei in situ is a very useful technique to evaluate embryo qualities and can be used as an indicator on study of preim-plantation embryo development.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.4
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• pp.339-348
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• 2010

Objective: To investigate the beneficial effect of fragment removal on the subsequent cell division and clinical outcome of the fragmented human embryos. Methods: A prospective study was performed in Hanna Women's Clinic and Mizmedi Hospital. Sixty couples undergoing In vitro fertilization-embryo transfer (IVF-ET) program were participated in the present study. The microsurgical fragment removal was performed in 106 fragmented embryos of 29 patients before the transfer. As a control group, 122 fragmented embryos of 31 patients were transferred without the fragment removal. Effects of fragment removal on morphological changes and clinical outcomes of fragmented embryos were investigated. Results: Mean morphological grade (G2.79) of fragmented embryos was significantly improved after the fragment removal(G1.63, p<0.001). Most of the fragmented embryos did not show a regeneration of fragments after the fragment removal during the subsequent development, and a beneficial effect of fragment removal on the development of the fragment removed embryos was observed. Implantation and pregnancy rates of fragment removed embryos were 12.3% and 31.3%, whereas the rates of control group embryos were 6.6% and 22.5%, respectively. There was no statistical significance in the rates between the two groups because of the low number of trials. Conclusion: Microsurgical fragment removal improved the subsequent development as well as the morphological grade of fragmented embryos. The fragment removal may be beneficial for neighboring blastomeres by repairing the intercellular communication and removing the secretion of the potential toxic materials by fragments.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.1
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• pp.65-72
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• 2010

Objective: To compare the respective pregnancy outcomes of cycles undergoing elective two cleavage-stage embryos transfer (2ET) and three cleavage-stage embryos transfer (3ET) in fresh in vitro fertilization and embryo transfer (IVF-ET) program. Methods: We conducted a retrospective matched case control study that included 100 women with 2ET and 100 women with 3ET from January 2007 to June 2009. Subjects were matched for reproductive profiles and cycle characteristics. All of transferred embryos in both groups had good qualities. Pregnancy rates (PR), implantation rate, and multiple PR were compared. Results: Demographics, stimulation parameters and embryological data were comparable in both groups. Main pregnancy outcomes with 2ET and 3ET groups were not statistically different; implantation rate (41.0% vs. 35.3%), positive pregnancy rate (58.0% vs. 60.0%), clinical PR (55.0% vs. 59.0%), ongoing PR (51.0% vs. 55.0%), respectively. However, the 3ET group showed significantly higher multiple pregnancy and triplet pregnancy rates (30.9% vs. 50.8%, p=0.031; 1.8% vs. 11.9%, p=0.036, respectively). Conclusion: In women with favorable conditions and good quality embryos undergoing IVF, 2ET can get pregnancy outcomes comparable to those of 3ET and reduce multiple pregnancy (especially, triplet pregnancy).