• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2002.11a
• /
• pp.73-73
• /
• 2002

Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.

• Journal of Embryo Transfer
• /
• v.11 no.2
• /
• pp.125-135
• /
• 1996

The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.3
• /
• pp.361-368
• /
• 1997

A total of 55 patients with various etiologies of infertility particitated in a study comparing two regimens of controlled ovarian hyperstimulation (COH) with GnRH agonists and gonadotropins. Nineteen patients were given an ultra-short stimulation protocol when the agonist was administered for 3 day from Day 2 of the cycle. The remaining 36 patients were given a long stimulation protocol when the agonist was administered from the mid-luteal phase of the cycle preceding the stimulation cycle. The mean number of gonadotropins used per patient was not different between two groups. No significant differences were found in the mean number of oocytes recovered, fertilization rate and embryo cleavage rate between two groups. Pregnancy and delivery rates were higher in ultra-short protocol than in long protocol, but these were not significant. These results suggest that an ultra-short protocol is as effective as a long protocol in in-vitro fertilization and embryo transfer.

• Journal of Embryo Transfer
• /
• v.17 no.2
• /
• pp.123-128
• /
• 2002

The present study aimed at determining the effective dose of Folltropin, a follicle timulating hormone (FSH), on superovulation in indigenous cows of Bangladesh. Fifteen regularly cycling 5～7 years old dry cows, weighing 200～250 kg with 2.5～3.0 body condition scores (BCS) were divided into three groups (n＝5). Individual groups were superovulated with 100, 200 or 300 mg of Folltropin per animal. The superovulation treatment was initiated at Day 10 or Day 11 of the estrous cycle (Day 0＝day of estrus). Alfaprostol (6 mg) was injected to each cow 72 h after the initiation of superovulation treatment to induce eestrus. After confirming standing estrus, the cows were inseminated 2～3 times, 12 h apart, depending on the duration of estrus. At Day 6 or Day 7, individual horns of the uterus were flushed with 150～200 $m\ell$ of phosphate buffered saline supplemented with BSA (0.2％), penicillin (100 IU／$m\ell$) and streptomycin (100 $\mu\textrm{g}$／$m\ell$) using a two-way foley catheter. The embryos were concentrated, removing the excess medium through an embryo filter, and identified under a stereomicroscope. The identified embryos were collected, washed four times, evaluated and graded as excellent, good, fair or poor. The excellent, good and fair embryos were considered as transferable quality embryos. The mean (range). numbers of embryos collected vs. transferable quality embryos far 100, 200 and 300 mg of Folltropin were 4.5 (1～10) vs. 3.5 (1～8); 2.5 (1～4) vs. 1 (0～2) and 0.0 (0～0) vs. 0.0 (0～0), respectively, Folltropin at a dose of 100 or 200 mg produced suitable ovarian stimulation for superovulation in indigenous zebu cows of Bangladesh. A dose of 300 mg or more Folltropin consistently caused preovulatory corpora lutea formation in the ovaries and resulted in zero embryo recovery.

• Journal of Embryo Transfer
• /
• v.17 no.1
• /
• pp.23-32
• /
• 2002

To establish an effective in vivo Hanwoo embryo production system, several factors including types of FSH analogue, FSH treatment day, grade of Corpus Luteum were examined. Among FSH analogues. Super-OV was shown to have lower effect on transferable embryo production (P<0.01). Regarding the day of FSH treatment, there were significant difference in estrus status of donors(P<0.1), fertilization rate, and transferable and freezable embryo production(P<0.05). Finally, embryo production was significantly(P<0.01) higher in grade A (8.06 ∼9.49) Corpus Luteum which showed more than 6 on the left or right ovary than in grade B, C and D(2.93∼6.28).

• Clinical and Experimental Reproductive Medicine
• /
• v.43 no.4
• /
• pp.233-239
• /
• 2016

Objective: Uterine contraction induced by the embryo transfer (ET) process has an adverse effect on embryo implantation. The aim of this study was to determine the effect of oxytocin antagonist supplementation on the day of ET on in vitro fertilization outcomes via a meta-analysis. Methods: We performed a meta-analysis of randomized controlled trials (RCTs). Four online databases (Embase, Medline, PubMed, and Cochrane Library) were searched through May 2015 for RCTs that investigated oxytocin antagonist supplementation on the day of ET. Studies were selected according to predefined inclusion criteria and meta-analyzed using RevMan 5.3. Only RCTs were included in this study. The main outcome measures were the clinical pregnancy rate, the implantation rate, and the miscarriage rate. Results: A total of 123 studies were reviewed and assessed for eligibility. Three RCTs, which included 1,020 patients, met the selection criteria. The implantation rate was significantly better in patients who underwent oxytocin antagonist infusion (19.8%) than in the control group (11.3%) (n = 681; odds ratio [OR], 1.92; 95% confidence interval [CI], 1.25-2.96). No significant difference was found between the two groups in the clinical pregnancy rate (n = 1,020; OR, 1.57; 95% CI, 0.92-2.67) or the miscarriage rate (n = 456; OR, 0.76; 95% CI, 0.44-1.33). Conclusion: The results of this meta-analysis of the currently available literature suggest that the administration of an oxytocin antagonist on the day of ET improves the implantation rate but not the clinical pregnancy rate or miscarriage rate. Additional, large-scale, prospective, randomized studies are necessary to confirm these findings.

• Journal of Embryo Transfer
• /
• v.23 no.3
• /
• pp.217-222
• /
• 2008

In previous studies, we reported that sow which was transferred OPS-freezing embryos not able to deliver a piglet (Kim et al, 2004). This study was conducted to investigate a possibility of gilt as recipients which produce piglets after transfer of OPS-freezing embryos. All transferred embryos were prepared by in vitro production (IVP) system. In vitro culture (IVC) medium used glucose-free NCSU23 supplemented with 5mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$. From day 3 of IVC, 10% fetal bovine serum albumin was added to the culture medium. In preparing of freezing embryos, embryos were treated with 7.5 $\mu g/ml$ cytochalasin-B for 30 min and centrifuged at $13,000{\times}g$ for 13 min. And then, embryos were exposed sequentially to an ethylene glycol (EG) solution, aspirated into open pulled straw (OPS), and plunged or thawed into the liquid nitrogen. In embryo transfer (ET), we used two kinds of type (surgical method vs. non-surgical method). In surgical method of embryo transfer, $55\sim65$ embryo were transferred in both uterine horn of two recipient gilts by plastic straw. Non-surgical method which is like artificial insemination was performed on three gilts. Each 140 frozen embryos were transferred to two gilts and 40 fresh embryos to one gilt. Pregnancy establishment was shown one recipient at 45 days after ET. However, the one recipient was also aborted at 58 days after ET. These results suggest that gilts can be considered as a candidate of recipients for OPS-freezing embryo transfer.

• Clinical and Experimental Reproductive Medicine
• /
• v.38 no.4
• /
• pp.186-192
• /
• 2011

Objective: Since IVF program was first established, various types of media and culture systems have been developed either in-house or commercially. The aim of this study was to compare the efficacy of in-house Maria Research Center (MRC) media to that of commercially available Sydney IVF media in human day 3 embryo transfer cycles. Methods: Three hundred sixty nine couples were included in this prospective, randomized, and comparative study. All couples undergoing IVF treatment at the Maria Fertility Hospital were randomly assigned to either Sydney IVF (n=178) or MRC (n=191) media. Results: No difference was observed between the MRC media and Sydney IVF media groups with respect to fertilization rate (74.4% vs. 75.5%). The clinical pregnancy and implantation rates of MRC media (47.1% and 20.0%, respectively) were also similar to those of Sydney IVF media (44.4% and 19.4%, respectively). However, the proportion of embryos with good quality on day 3 was significantly higher in the MRC media group than the Sydney IVF media group (50.2% vs. 43.2%) ($p$ <0.05). Conclusion: MRC media were as effective as Sydney IVF media for sustaining embryo development and pregnancy rates. The present study implies that MRC media can be a suitable alternative to commercially available media for human IVF-ET program.

• Clinical and Experimental Reproductive Medicine
• /
• v.36 no.4
• /
• pp.293-300
• /
• 2009

Objective: This study was aimed to investigate whether uterine blood flow on the day of embryo transfer can be a predictor of pregnancy outcome in human IVF. Methods: Fifty-one patients undergoing IVF program were included in this study. Serum estradiol levels were measured on the day of hCG administration and uterine pulsatility index (PI) and resistance index (RI) was examined for at embryo transfer of day 3. Results: Of 51 cycles, 22 cycles were clinically pregnant (43.1%) and the implantation rate was 14.7%. Uterine PI and RI had a significant inverse correlation with serum estradiol levels (p<0.05). These uterine blood flows were not significantly different between pregnant and nonpregnant groups. The pregnancy rate was slightly higher in patients with PI more than 3.0 compared to those with PI of 3.0 or less, but there was no significant difference. Conclusion: These results suggest that uterine PI and RI at the day of embryo transfer could not be a good predictor of pregnancy in IVF treatment. But they had an inverse correlation with serum estradiol levels on the day of hCG administration.

• Journal of Embryo Transfer
• /
• v.12 no.3
• /
• pp.301-306
• /
• 1997

The present study was carried out to investigate effect of FSH -P dose and energy level on normal embryo production after superovulation in Hanwoo. The results obtained were as follows ; 1. There was a significant effect of dose of FSH-P on normal embryo production in Hanwoo(P$\pm$5.9), 40(4.9i5.7), 50mg(2.2$\pm$2.6). 2. The plasma P$_4$ levels on the first treatment day were higher group( >4ng /ml) than lower group( <=4ng /ml), produced significicantly(P<0.05) higher number of normal embryos. 3. There was a significant effect of energy level on normal embryo production in Hanwoo(P$\pm$6.0), number of normal embryos were higher than TDN 70%(5.1$\pm$6.5) and TDN 130%(4.4$\pm$2.6) 4. The donor returned to normal estrus after superovulation were 44.8, 28.4 and 29.9 days by TDN 70, 100 and 130%, respectively.