• Title/Summary/Keyword: In vitro fertilization and embryo transfer (IVF-ET)

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Production of Transgenic Cattle by Non-surgical Embryo Transfer (비외과적 수정란 이식에 의한 형질전환 소 생산 기술)

  • Uhm, Sang Jun;Yang, Jung Seok;Lee, Su Min;Joe, So Young;Heo, Young-Tae;Xu, Yong-Nan;Koo, Bon Chul;Cheong, Ki Soo;Kim, Kwang Jae;Kim, Ji Tae;Kim, Nam-Hyung;Ko, Dae-Hwan
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.169-175
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    • 2013
  • Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{\mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{\mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

Influence of Maternal Age on Embryo Quality and the Frequency of Multiple Pregnancy in IVF-ET Program (시험관 아기 시술에서 여성의 연령이 수정란의 질과 다태 임신 발생에 미치는 영향)

  • Lee, Myeong-Seop;Park, Jang-Ok;Jung, Ji-Hak;Park, Jun-Suk;Kang, Hee-Gyoo;Kim, Dong-Hoon;Lee, Ho-Joon
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.261-265
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    • 2000
  • Objective: This study was performed to evaluate the influence of maternal age on embryo quality and the frequency of multiple pregnancy in IVF-ET program. Method: 86 conventional IVF-ET cycles were divided into three groups according to the age by 5 year (group A: 26-30, group B: 31-35, group C: 36-40 yrs). The in vitro fertilization and development outcome (fertilization, cleavage and high quality embryo rate) and the pregnancy outcome (pregnancy, implantation, G-sac/high quality embryo and multiple pregnancy rate) were examined. And then, these results were compared among the groups. Results: The rates of fertilization (62.7, 68.5 and 65.4%, respectively) and cleavage (95.6, 97.6 and 98.0%, respectively) were not different among the groups. And the high quality embryo (HQE) rate also was not different among the groups (61.8, 62.9 and 62.8%, respectively). The pregnancy rate of group C (23.3%) was significantly lower than that of group A (41.2%) and B (48.7%). And the implantation rate was significantly decreased with advance in maternal age (group A; 17.3%, B; 12.6% and C; 6.0%). The G-sac/high quality embryo rate was significantly higher in group A (70.8%) when compared to group B (32.2%) and C (40.0%). On the other hand, the multiple pregnancy rate was significantly lower in group C (14.3%) when compared to group A (71.4%) and B (36.8%). Conclusion: The pregnancy rate was significantly decreased over 35 years. The G-sac/HQE and multiple pregnancy rate were significantly high below 31 years. Thus, these results suggest that the number of high quality embryo transferred should be limited by the age and another criteria for embryo quality evaluation were required for single embryo transfer.

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Sex Determination of Biopsied Hanwoo Embryos by Polymerase Chain Reaction and Embryo Transfer with Sexed Blastocysts (한우 체외수정란 Biopsy 후 PCR 기법을 이용한 성 판정과 성감별 수정란의 이식)

  • 김용준;정구남;이해이;조성우;김용수;유일정
    • Journal of Embryo Transfer
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    • v.15 no.3
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    • pp.219-230
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    • 2000
  • This study was carried out to determine the factors on achieving good viability of embryos biopsied fur sexing, to investigate pregnancy rate following embryo transfer(ET) with sexed embryos, and to confirm the accuracy for the calves bort following ET with sexed embryos by polymerase chain reaction(PCR). To investigate viability of Hanwoo embryos after biopsy for sexing, fresh and frozen/thawed embryos were biopsied according to different developmental day of blastocysts, different stage of blastocysts, and different biopsy grade and the embryos themselves were incubated for 2 hours in TCM199 after microsection to be evaluated morphologically for recovery as blastocyst. The results obtained were as follows : 1. The rate of oocytes cleaved in vitro and the rate of blastocyst of the cleaved oocytes were 52.5% and 21.6%, respectively. The rate of blastocyst on day 8 was 11.2%, denoting the highest rate during whole culture period posterior to in vitro fertilization(IVF) 2. After biopsy for sexing, the viability rate of blastocyst on day 7, 8 and 9 was 75.0%, 88.4%, and 100.0%, respectively and the viability of early, mid, and expanded blastocyst after biopsy was 75.0%, 88.9%, and 91.1%, respectively The viability rate of fresh and frozen/thawed embryos was 89.9%, 71.4%, respectively. And the viability of expanded, hatching, and hatched blastocyst of frozen/thawed embryos was : 75.0%, 75.0%, and 50.0%, respectively. The viability of embryos according to biopsy grade of 10∼20%, 21∼30%, and 31∼40% was 85.7%, 91.5%, and 71.4%, respectively. 3. Pregnancy rate after transfer with biopsied embryo between flesh and frozen/thawed embryos was 22.6% and 20.0%, respectively. 4. In comparison between sex by PCR method and sex of calves born after embryo transfer, the accuracy of sex deterimination was 92.3% (12/13).

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A Case of Primary Abdominal Pregnancy on Uterosacral Ligament Following in vitro Fertilization and Embryo Transfer (시험관아기 시술 후 자궁천골인대에 발생한 일차성 복강임신 1예)

  • Won, Hyung-Jae;Yoon, Tae-Ki;Seok, Hyun-Ha;Kim, Yu-Sin;Lee, Kyung-Ju;Kwon, Tae-Hee
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.2
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    • pp.133-137
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    • 2007
  • Abdominal pregnancy is a very rare form of ectopic pregnancy following IVF-ET procedure. Because the symptoms of abdominal pregnancy is not typical, it is difficult to diagnose early and the mortality rate is higher than other ectopic pregnancies. Liver, spleen, omentum and diaphragm are main implantation sites of abdominal pregnancy. We reported an unusual first case of an etopic pregnancy on uterosacral ligament following in vitro fertilization and embryo transfer with brief review of literature.

Application of Embryo Transfer Technology (수정란 이식 기술의 응용)

  • Lim, Hyun-Joo;Son, Jun-Kyu;Yoon, Ho-Beak;Baek, Kwang-Soo;Choe, Chang-Yong;Kim, Sidong;Kwon, Eung-Gi
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.163-168
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    • 2013
  • Embryo transfer (ET) technology is of high importance in modern cattle breeding programs. ET is one step in the process of removing one or more embryos from the reproductive tract of an outstanding donor female and transferring them to one or more recipient females. Embryos also can be produced in the laboratory via techniques such as in vitro fertilization (IVF). But the actual transfer of an embryo is only one step in a series of processes that may include some or all of the following: superovulation and insemination of donors, collection of embryos, isolation, evaluation and short-term storage of embryos, micromanipulation and genetic testing of embryos, freezing of embryos and embryo transfer. Cryopreservation and direct transfer of frozen-thawed embryos is common-place with pregnancy rates near that of fresh embryos. Polymerase chain reaction (PCR) technology is currently being used for sexing embryos, and this technology will be used for "embryo diagnostics" and "embryo genomics" in the future. Although, many limitations and problems remain to overcome, these and other new technologies promise to change livestock breeding drastically in the next decade.

The Effectiveness of Moxibustion Treatment in Infertility with IVF-ET: A Systematic Review and Meta-Analysis (보조생식술을 시행한 난임환자에서의 뜸 치료에 대한 체계적 문헌고찰 및 메타분석)

  • Lee, Ho-Sung;Park, Yong-Duk;Lee, Hye-Jung;Hwang, Deok-Sang;Jang, Jun-Bock;Lee, Chang-Hoon;Lee, Jin-Moo;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.35 no.2
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    • pp.28-41
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    • 2022
  • Objectives: The purpose of this study is to investigate the effectiveness of moxibustion in infertility with In Vitro Fertilization and Embryo Transfer (IVF-ET). Methods: We searched 8 databases (Embase, PubMed, CiNii, CNKI, OASIS, ScienceOn, KMBASE, KISS)to identify eligible studies published before 2021 Oct. We included randomized controlled clinical trials (RCTs) using moxibustion in infertility with IVF-ET. The methodological quality of each RCT was assessed by the Cochrane risk of bias tool. Results: Two RCT studies were eligible in our review. The overall risk of bias was evaluated as unclear. The meta-analysis of 2 trials indicated that favorable results for the use of moxibustion with IVF-ET. Conclusions: This systematic review and meta-analysis of clinical trials suggests that moxibustion with IVF-ET can effect on Infertility patients. However, because of studies included analysis was biased due to unclear risk of bias and unreliable study design, future RCT studies and additional Meta-Analysis are needed to judge the supplementary treatment role of moxibustion in infertility with IVF-ET.

Improvement of Pregnancy Rates by Coculture of Human Embryos with Cumulus Cells in Glucose and Phosphate Free M-TALP Media (Glucose와 Phosphate가 제거된 M-TALP 배지에서의 난구세포 공배양에 의한 임신율 향상에 관한 연구)

  • Chung, B.S.;Chang, W.H.;Lee, M.H.;Kim, J.Y.;Bang, J.H.;Kim, K.H.;Suh, T.K.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.75-81
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    • 1999
  • The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

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Mouse Embryo Culture as Quality Control for Human IVF:Culture Media and Supplements (인간 난자의 체외수정을 위한 정도관리로서 생쥐 착상전 배아의 배양에 관한 연구)

  • Lee, Gy-Soog;Park, Jong-Duk;Lee, Choon-Khoon;Kim, Jong-Duk
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.2
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    • pp.161-171
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    • 1989
  • The development of 2-cell mouse embryos to the blastocyst stage in vitro has been used as quality control for the culture media and supplements employed for human in vitro fertilization and embryo transfer(IVF-ET). 2-cell mouse embryos were cultured to the blastocyst stage in SECM, Medium 199-Earle's, Ham's F-10 I , Ham's F-10 II , Hoppe & Pitts, MEM and $HT_6$. The protein supplements contained in media were bovine serum albumine, fetal bovine serum and human fetal cord serum. The results were as follows; 1. The successful development was 81.3% in Medium 199-Earle’s, 91.9% in Ham’s F-10 I and 97.1% in $HT_6$. 2. 2-cell mouse embryos developed properly in all supplements but the best development was particularly noted in $HT_6$ media when HFCS was supplied as protein supplement.

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Effect of a Baseline Ovarian Cyst on the Outcome of in Vitro Fertilization-embryo Transfer (과배란유도 전 기저 난소 낭종이 체외수정시술 결과에 미치는 영향)

  • Bai, S.W.;Lee, K.J.;Lee, B.S.;Park, K.H.;Cho, D.J.;Song, C.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.179-183
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    • 1999
  • This study was performed to determine the significance of a baseline ovarian cyst on the response to controlled ovarian hyperstimulation and the outcome of IVF-ET. One hundred one patients who underwent IVF-ET were enrolled in this study. The outcome of 31 patients, who had an ovarian cyst of >10mm detected at ultrasound examination performed on day 3, was compared with that of 70 patients who underwent a similar protocol and did not have an ovarian cyst. E2 level on the day of hCG administration, the number of follicles, the number of oocytes retrieved, the number of embryo transferred and the pregnancy rate were evaulated. The E2 level on the day of hCG adminstration and the number of mature oocytes retrieved were lower in the group with a baseline cyst. The pregnancy rate also was significantly lower in the group with a cyst (21% versus 38%). Therefore a baseline ovarian cyst on cycle day 3 was associated with a poorer outcome after IVF-ET.

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Luteal Phase Serum Estradiol and Progesterone in In Vitro Fertilization and Embryo Transfer (인간난자의 체외수정 및 배아의 자궁내이식 후 황체기 혈청 난포호르몬 및 황체호르몬의 변동에 관한 연구)

  • Lee, Jin-Yong;Kim, Hak-Soon;Kim, Jung-Gu;Moon, Shin-Yong;Chang, Yoon-Seok
    • Clinical and Experimental Reproductive Medicine
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    • v.14 no.2
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    • pp.127-137
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    • 1987
  • Steroid hormone profiles during luteal phases after in vitro fertilization(IVF) and embryo transfer(ET) have been evaluated in 83 cycles stimulated by pure follicle-stimulating hormone/human menopausal gonadotropin/human chorionic gonadotropin, in which 13 patients became pregnant. Serum estradiol($E_2$) and progesterone($P_4$) levels were determined on days 2, 5, 7 and 9 after laparoscopic follicle aspiration. The follicular $E_2$ peak was slightly higher in pregnancies than in failures. Positive correlations were observed between the follicular $E_2$ peaks and the $P_4$ levels on days 5 and 7 of the luteal phase in pregnancies, but no correlation was found in failures. The $E_2$ and $P_4$ levels on days 5 and 7 of the luteal phase were significantly higher in pregnancies than in failures, but not different on days 2 and 9. Values of the $P_4/E_2$ ratio were similar between the two groups. The luteal phase durations were 12 to 19 days and no correlation was observed between the lengths of luteal phase and the luteal $E_2$ or $P_4$ concentrations. These data suggest that high $P_4$ levels in the mid-luteal phase, which have positive correlations with the follicular $E_2$ peaks, might have a favorable influence on the pregnancy success in human IVF.

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