• Title/Summary/Keyword: oocyte transfer

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Effects of Donor Cells and Estrus Synchronization on the Production of Cloned Korean Native Goat (공핵 세포 및 발정 동기화가 복제 재래 산양 생산에 미치는 영향)

  • Park H.S.;Kim T.S.;Jung S.Y.;Park J.K.;Lee J.S.;Jung J.Y.
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.137-146
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    • 2006
  • The objective of this study was to examine the effect of donor cell types, the source of recipient oocytes and estrous synchronization on pregnancy and delivery rates of somatic cell nuclear transfer (SCNT) embryos in Korean native goats. Recipient oocytes were surgically collected after superovulation. Ear cells and fetal fibroblasts were collected and cultured in serum-starvation condition (TCM-199 + 0.5% FBS) for cell confluence. The zonae pellucidae of in vivo- and in vitro-matured oocytes were partially drilled using a laser system. Single somatic cell was transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3 M mannitol. After the fusion, embryos were activated by Ionomycin+6-DMAP. NT embryos were cultured in mSOF medium supplemented with 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 12 to 20 hr. One hundred and two SCNT embryos were transferred into 20 recipients and pregnancy rate at days 30 was 20.0%. Of them, one developed to term and delivered 1 kid. Ear cells showed significantly higher fusion (63.8 vs. 26.5%) and pregnancy rates (20.0 vs. 0.0%) than those of fetal fibroblast (p<0.05). The recipients synchronized by CIDR showed significantly lower pregnancy rates compared to that of recipient in natural estrus ($0.0{\sim}25.0%$ vs. 100%) (p<0.05). Cloned kid was born from the recipient in natural estrus. For the synchronization of estrus between recipient and donor, there was no difference between treatments (${\pm}0$ vs. +12 hr) in pregnancy rate. The first healthy cloned kid (Jinsoonny) was produced by transfer of SCNT embryos derived from in vivo oocytes and ear cells into a recipient goat whose estrus was synchronized with the donor. These results imply that donor cells for nuclear transfer may affect the success rate, and the estrus synchronization between donor and recipient animals can also be important.

Heat waves impair cytoplasmic maturation of oocytes and preimplantation development in Korean native cattle (Hanwoo)

  • Sa, Soo Jin;Jeong, Jiyeon;Cho, Jaesung;Lee, Seung-Hwan;Choi, Inchul
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.493-498
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    • 2018
  • There has been widespread warming and a general increase in summer temperatures over the Korean peninsula ($0.5^{\circ}C$/10 years from 2001 to 2010). South Korea is transforming into a subtropical region, and the productivity of livestock is affected by the climatic changes. In this study, we investigated whether the summer heat waves affect the developmental competency of Korean native cattle (Hanwoo), a taurine type of cattle with a small portion of indicine varieties. We collected oocytes during the summer (heat stress, HS) and autumn (non-HS condition) and examined the developmental competencies including in vitro maturation and preimplantation embryo development. No significant differences were observed between the HS and non-HS oocytes in nuclear maturation (extrusion of the polar body); however, the cleavage and blastocyst rates were significantly lower in the HS group than those in the non-HS group. The lower developmental competence of the HS oocytes compared to the non-HS is, in part, due to insufficient cytoplasmic maturation because of a higher production of Reactive oxygen species (ROS) levels as well as peri/cortical distributed mitochondria in the HS oocytes after in vitro maturation. Next, we examined the ROS and mitochondria distribution and found a significant increase in the levels of ROS in the HS oocytes and a polarized distribution (pericortical cytoplasm) of mitochondria in the HS oocytes. In summary, impaired cytoplasmic maturation of oocytes from exposure to HS affects the preimplantation embryo development by dysfunction of mitochondria. To improve reproductive performance, embryo transfer using cryopreserved embryos/oocytes is recommended in the hot summer season of South Korea.

A Study for GnRH Antagonist (Cetrotide) Short Protocol in Controlled Ovarian Hyperstimulation (GnRH Antagonist (Cetrotide) Short Protocol의 임상적 유용성에 관한 연구)

  • Kim, Moon-Young;Jung, Byeong-Jun
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.4
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    • pp.265-270
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    • 2001
  • Objective : The aim of this study was to evaluate the outcome the GnRH antagonist (Cetrotide) short protocol in controlled ovarian hyperstimulation comparing with GnRH agonist long protocol. Materials and Method: From July 2000 to November 2001, 26 patients, 28 cycles were performed in controlled ovarian hyperstimulation by GnRH antagonist and GnRH agonist. GnRH antagonist (Cetrotide) was administered in 12 patients (14 cycles, Group 1) and GnRH agonist (Lucrin, Sub Q, Group 2) in 14 patients (14 cycles). Ovulation induction was performed by hMG (Pergonal) in group 1, and by Combo (Metrodine HP + Pergonal) in group 2. We compared the fertilization rate, good quality embryo, and clinical pregnancy rate between the two groups. Student-t test and Chi-square were used to determine statistical significance. Statistical significance was defined as p<0.05. Results: Ovarian hyperstimulation syndrome did not occurred in which estradiol (E2) level was $3874{\pm}809\;pg/ml$ and the number of retrieved oocytes was $18.4{\pm}2.4$. The number of used gonadotropin ampules was significantly decreased in Group 1 (26.0 vs. 33.1, p<0.04). There were no significant difference in the number of preovulatory oocyte ($10.6{\pm}6.9$ vs. $10.0{\pm}6.1$), fertilization rate ($74.8{\pm}23.4$ vs. $72.2{\pm}21.8$), good quality embryo ($58.7{\pm}23.6$ vs. $38.7{\pm}36.6$), and embryo transfer ($4.3{\pm}1.6$ vs. $4.4{\pm}1.6$). Although the age of the group 1 was older than the group 2 (34.4 vs. 30.8), there was no significant difference in clinical pregnancy rate (50.0% vs. 57.1%). Conclusions: We suggest that GnRH antagonist was a safe, effective, and alternative method in the controlled ovarian hyperstimulation, especially in PCOD patients who will be develop the ovarian hyperstimulation syndrome.

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Correlation between Ultra sonic Preovulatory Follicular Appearances and the Retrieval of Mature Oocytes in Stimulated Cycles (과배란유도후 배란직전 난포의 초음파검사 소견과 성숙난자 획득간의 상관관계에 관한 연구)

  • Kim, Hak-Soon;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Lee, Jin-Yong;Chang, Yoon-Seok
    • Clinical and Experimental Reproductive Medicine
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    • v.15 no.1
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    • pp.25-34
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    • 1988
  • The intrafo1licular echoes of cumulus oophoruses within ovarian follicles were assessed with the use of ultrasound in 86 women taking part in an in vitro fertilization(IVF) or gamete intrafallopian transfer(GIFT) program, stimulated with pure follicle-stimulating hormone(FSH)/human menopausal gonadotropin(hMG)/human chorionic gonadotropin (hCG). When intrafo1licular echoes were clearly separated from the follicular wall or relatively dispersed within the follicle, they were considered to be a dissociated cumulus, and when they were only slightly prominent from the follicular wall, they were suspected to be a nondissociated cumulus. A cumulus was seen in 62.1% of the follicles larger than 10 mm diameter and 75.1% of them were dissociated. The larger the follicles in size, the more the cumuluses in number and dissociation. The number of follicles and intrafollicular echoes per woman was not different whether or not she would be pregnant, but the number of dissociated cumuluses was significantly more in pregnant women. The number of observed dissociated cumuluses correlated significantly with the number of recovered mature oocytes. When an intrafollicular echo is seen, it can be taken as evidence of a sign of maturity of that particular follicle and oocyte. Ultrasonographic monitoring of intrafollicular echoes and follicular size is very helpful to predict follicular maturation in ovulation stimulation cycles.

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Live birth outcomes of vitrified embryos generated under growth hormone stimulation are improved for women categorized as poor-prognosis

  • Keane, Kevin N;Ye, Yun;Hinchliffe, Peter M;Regan, Sheena LP;Dhaliwal, Satvinder S;Yovich, John L
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.4
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    • pp.178-188
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    • 2019
  • Objective: To determine the clinical pregnancy (CP) and live birth (LB) rates arising from frozen embryo transfers (FETs) that had been generated under the influence of in vitro fertilization (IVF) adjuvants given to women categorized as poor-prognosis. Methods: A registered, single-center, retrospective study. A total of 1,119 patients with first FETs cycle include 310 patients with poor prognosis (109 treated with growth hormone [GH], (+)GH group vs. 201 treated with dehydroepiandrosterone, (-)GH group) and 809 patients with good prognosis (as control, (-)Adj (Good) group). Results: The poor-prognosis women were significantly older, with a lower ovarian reserve than the (-)Adj (Good) group, and demonstrated lower chances of CP (p< 0.005) and LB (p< 0.005). After adjusting for confounders, the chances of both CP and LB in the (+)GH group were not significantly different from those in the (-)Adj (Good) group, indicating that the poor-prognosis patients given GH had similar outcomes to those with a good prognosis. Furthermore, the likelihood of LB was significantly higher for poor-prognosis women given GH than for those who did not receive GH (p< 0.028). This was further confirmed in age-matched analyses. Conclusion: The embryos cryopreserved from fresh IVF cycles in which adjuvant GH had been administered to women classified as poor-prognosis showed a significant 2.7-fold higher LB rate in subsequent FET cycles than a matched poor-prognosis group. The women with a poor prognosis who were treated with GH had LB outcomes equivalent to those with a good prognosis. We therefore postulate that GH improves some aspect of oocyte quality that confers improved competency for implantation.

Acupuncture as an adjunct treatment to increase the success rate of in vitro fertilisation: an overview of systematic reviews and meta-analyses

  • Jeong, Daun;Hyun, Min-Kyoung;Jeong, Jae-Cheol;Park, Jang-Kyung;Choi, Min-Sun;Kim, Dong-Il;Lee, Dong-Nyung
    • The Journal of Korean Medicine
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    • v.35 no.2
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    • pp.1-11
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    • 2014
  • Objectives: To evaluate the effect of acupuncture as an adjunct treatment to increase the success rate of in vitro fertilisation (IVF). Methods: A review of the English and Korean literature was conducted to identify studies on acupuncture as an adjunct treatment to IVF. The main outcome measures were the biochemical pregnancy rate (BPR), the implantation rate (IR), the clinical pregnancy rate (CPR), the miscarriage rate (MR), the on-going pregnancy rate (OPR) and the live birth rate (LBR). Results: Nine meta-analyses of 11 systematic reviews (SRs) were included in this review. In four SRs, the overall IVF outcomes regardless of the procedural steps were documented; two of these SRs reported a significant effect on the CPR. One SR reported that acupuncture at the time of controlled ovarian hyperstimulation had a significant effect on the BPR. Seven SRs reported that acupuncture had no significant impact on transvaginal oocyte retrieval (TVOR). In eight SRs, significant impacts on the CPR, LIBR and OPR were reported when acupuncture was performed around the time of embryo transfer (ET). The results of repeated acupuncture after ET were included in two SRs, which included the same primary studies. The results of the SRs showed that acupuncture had a positive effect on the CPR. Conclusions: When the complete IVF procedure is analysed, there is conflicting evidence in that some reviews have found that acupuncture leads to an increased CPR but others have not. In addition, the reviews presented no evidence to suggest that acupuncture has any specific risks.

Studies on Production and Efficient Utilization of Livestock Embryos by In Vitro Fertilization and Miceomanipulation I. Transfer and Freezing of In Vitro Fertilized Rabbit and Bovine Oocytes Matured In Vitro (체외수정 및 미세조작에 의한 가축배의 생산가 효율적 이용에 관한 연구 I. 체외성숙.체외수정된 토끼 및 소 배의 이식과 동결)

  • 김창근;정영채;이근상;김희석;정진태;윤종택;최미호;이장희;김광식
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.57-68
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    • 1993
  • This stduy was carried out to find a reliable method for the production of in vitro fertilized embryos having more excellent development capacity and freezability in the rabbit and cattle. The greatest number of rabbit oocytes was recovered 6hrs after HCG injection(P<0.05). The maturation rate in vitro was slightly higher in the oocytes(6-h-oocytes) from 6h than those (8-h-oocytes)from 8 hrs after HCG injection and the beneficial effect of FSH during oocyte maturation was significantly great in the oocytes from large follicles. The cleavage rate into 2-to-6-cell stage was not differ between the 6-h-oocytes and 8h-oocytes, but the cleavage of these oocytes was greatly promoted by FSH addition to maturation medium and the cleavge of 8-h-oocytes matured without FSH was significantly low. The embryo development into 16-cell to morula was not promoted by the co-culture with rabbit oviduct epithelial cells. The freezability by embryo stages was ovidusly high at 4-cell and morula stage in 6-h-oocytes and the viability of 16-cell embryos from 8h-oocytes was similar to that of morula stage. The implantation sites after surgical tranfer of fresh rabbit embryos were not implanted. In bovine experiment, the in vitro development into 16-cell and morula after in vitro maturation and fertilization in the follicular oocytes was slightly improved by the co-culture with granulosa cells compared to that with oviduct epithelial cells and the frozen-thawed viability rate of these embryos ranged from 14 to 40%. The excellent fresh embryos were transferred nonsurgically to 6 recipients, but were not pregnant.

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Pretreatment of normal responders in fresh in vitro fertilization cycles: A comparison of transdermal estradiol and oral contraceptive pills

  • Pereira, Nigel;Petrini, Allison C.;Zhou, Zhen N.;Lekovich, Jovana P.;Kligman, Isaac;Rosenwaks, Zev
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.4
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    • pp.228-232
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    • 2016
  • Objective: The aim of this study was to investigate the impact of pretreatment with transdermal estradiol ($E_2$) compared to oral contraceptive pills (OCPs) on controlled ovarian stimulation (COS) response in normal responders undergoing fresh in vitro fertilization (IVF)-embryo transfer (ET) cycles. Methods: A retrospective cohort study was performed of normal responders undergoing fresh IVF-ET cycles who received pretreatment with transdermal $E_2$ versus OCPs prior to fresh IVF-ET. The total days of ovarian stimulation, total dosage of gonadotropins, total number of oocytes, and mature oocytes retrieved were noted. Pregnancy outcomes after ET were also recorded. Results: A total of 2,092 patients met the inclusion criteria: 1,057 and 1,035 patients in the transdermal $E_2$ and OCP groups, respectively. Patients in the OCP group had a longer duration of COS ($10.7{\pm}1.63days$, p< 0.01) than the $E_2$ group ($9.92{\pm}1.94days$). Patients in the OCP group also required higher cumulative doses of gonadotropins ($2,657.3{\pm}1,187.9IU$) than those in the $E_2$ group ($2,550.1{\pm}1,270.2IU$, p= 0.002). No statistically significant differences were found in the total and mature oocytes retrieved or in the rates of biochemical pregnancy, clinical pregnancy, spontaneous miscarriage, and live birth between the groups. Conclusion: Our findings suggest that compared to OCPs, pretreatment with transdermal $E_2$ is associated with a shorter duration of ovarian stimulation and lower gonadotropin utilization, without compromising the oocyte yield or pregnancy outcomes in normal-responder patients undergoing fresh IVF.

Studies on Serum Estradiol (E2), Androstenedione (ADD) and Testosterone (T) Levels in the Hyperstimulated Ovulatory Cycles (과배란유도 월경주기에서 혈중 Estradiol Andrstenedione 및 Testerone농도의 동태에 관한 연구)

  • Kim, Jong-Hwa;Lee, Jin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.13 no.2
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    • pp.101-112
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    • 1986
  • It is now common practice to attempt ovarian hyperstimulation in vitro fertilization and embryo transfer (IVF-ET) to promote the development of multiple preovulatory follicles and to maximize the number of mature egg available. There are several drugs for hyperstimulation such as clomiphene citrate only, clomiphene citrate and human menopausal gonadotropin (HMG) and HMG only. Accumlated experience has shown that the hyperstimulation of the ovary in IVF-ET results in high pregnancy rate. But the hyperstimulation of the ovary in IVF-ET may cause the hyperandrogenism, so we must consider the adverse effect on pregnancy rate of the hyperandrogenism. Little is known about the functional significance of androgen for the follicular growth, however, the hyperandrogenism might interfere with oocyte maturation. The aim of the present investigation was to determine the serum profiles of estradiol, androstenedione and testosterone during the hyperstimulated menstrual cycles in IVF. The results were summarized as follows: 1. There was a gradual increase in the mean levels of serum estradiol, androstenedione, and testosterone approaching follicular maturation. 2. The mean serum estradiol levels in the hyperstimulated groups were significantly higher than that in the control group in late follicular phase and ovum retrieval (ovulation) day (p<0.01). 3. The mean serum androstenedione levels in the clomiphene citrate groups were significantly higher than that in the control group in late follicular phase (p<0.01). There was no statistically significant different in the mean serum androstenedione levels between the control group and the HMG group (p>0.05). 4. There was no statistically significant difference in the mean levels of testosterone among each group (p>0.05). 5. There was no statistically significant different in the mean levels of estradiol, androstenedione and testosterone between the fertilized patients and non-fertilized patients in clomiphene citrate and HMG group (p>0.05).

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Efficiency of laser-assisted intracytoplasmic sperm injection in a human assisted reproductive techniques program

  • Choi, Kyoung-Hee;Lee, Jung-Hyun;Yang, Yun-Hee;Yoon, Tae-Ki;Lee, Dong-Ryul;Lee, Woo-Sik
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.3
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    • pp.148-152
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    • 2011
  • Objective: Laser-assisted intracytoplasmic sperm injection (LA-ICSI), also known as micro-opening or thinning of the zona pellucida (ZP) prior to ICSI, may help to reduce mechanical damage to the oocyte during the procedure. The aim of the present study was to evaluate and analyze the efficacy of our institutional LA-ICSI program, which features laser-assisted ZP thinning prior to ICSI, in comparison with conventional ICSI (C-ICSI), performed on patients with different clinical characteristics. Methods: Patients undergoing a total of 212 ICSI cycles were randomly divided into an LA-ICSI group (106 cycles) and a conventional ICSI group (106 cycles). To reduce tissue damage, we thinned the ZP by approximately 70%, using a laser, before ICSI. Patients thus treated formed the LAICSI group. Comparisons included the morphological quality of transferred embryos, blastocyst development of the remaining embryos, and clinical pregnancy, in terms of ICSI method and patient characteristics. Results: Fertilization, development of remaining embryos, and pregnancy rate were significantly higher in the LA-ICSI group compared with the C-ICSI group. Fertilization, embryonic development, and the pregnancy rate were all improved in younger patients (<38 years of age) and in those who underwent a low number of IVF-ET attempts (<3 trials). In addition, the pregnancy rate was increased in older patients. Conclusion: LA-ICSI may be useful in improving the chance of pregnancy in all ICSI patients.