• Clinical and Experimental Reproductive Medicine
• /
• v.33 no.3
• /
• pp.149-157
• /
• 2006

Objective: This study was to investigate the clinical efficiency of clomiphene citrate/GnRH antagonist protocol comparing with the clomiphene citrate only protocol in infertile women with normal ovulatory cycles. Method: Among 116 patients, 43 were received assisted reproductive technologies using natural ovulatory cycle, 38 and 35 were received clomiphene citrate only protocol and clomiphene citrate/GnRH antagonist combined protocol, respectively, and the clinical results were compared and analyzed Results: In each group, basal levels of LH, FSH, $E_2$ and FSH, $E_2$ on hCG day injected were not different, but LH level and endometrial thickness on hCG injected day were decreased significantly and the pregnancy rate was increased significantly in clomiphene citrate/GnRH antagonist group. Conclusion: The pregnancy rate was increased significantly in clomiphene citrate/GnRH antagonist group compared with natural ovulatory cycle and clomiphene citrate only group.

• Journal of Embryo Transfer
• /
• v.26 no.3
• /
• pp.147-152
• /
• 2011

This study was conducted to examine whether efficiency of oocyte production from superovulated prepubertal goats. Fifteen prepubertal and twenty adult goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotrophin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes was activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at 38.5$^{\circ}C$ in an atmosphere of 5% CO$_2$, 5% O$_2$, 90% N$_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I + II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytcs. The cleavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes.

• Journal of Embryo Transfer
• /
• v.28 no.1
• /
• pp.19-24
• /
• 2013

This study assesses of efficiency of oocyte recovery and in vitro development for during the non breeding season in goat. Thirty-four matured goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injections of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotropin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes were activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I+II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytes. The clavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes. There was no significant difference in oocyte quality between seasonal treatments.

• Korean Journal of Animal Reproduction
• /
• v.27 no.3
• /
• pp.241-248
• /
• 2003

This study was conducted to determine the effects of progestagen (Synchromate-B, Veramix, CIDR) and hormone (PMSG, FSH, hCG) treatments on superovulation and estrous synchronization in Korean native goats. Goats were initially superovulated by using progestagens (Synchromate-B, Veramix, CIDR) and hormones (FSH, PMSG, hCG). The progestagens were removed after 15 days of insertion and 0.88 mg of FSH was injected intramuscularly twice a day from day 12 to day 15. In addition, 150 IU of PMSG and 200 IU hCG were injected intramuscularly in the morning of day 12 and in the afternoon of day 15, respectively. Estrous synchronization was induced by the progestagen releasing devices for 13 days and intramuscular injection of 400 IU PMSG in the morning of day 11. The results were summarized as follows: 1. The responses of superovulation treated with three types (Synchromate-B, Veramix, CIDR) of progestagen were 98.6, 99.4 and 98.8%, respectively. The average ovulation rates with Synchromate-B, Veramix and CIDR were 12.58 $\pm$ 6.52, 12.91 $\pm$ 7.27 and 11.28 $\pm$ 6.33, respectively. The average ovulation rates of Synchromate-B and Veramix treatments were significantly higher than that of the CIDR treatment (P<0.05). 2. The responses of estrus synchronization treated with Synchromate-B, Veramix and CIDR were 52.9, 72.9 and 75.7%, respectively. Veramix and CIDR treatments on estrous synchronization were significantly higher than the Synchromate-B treatment (P<0.05). Among the estrous synchronized goats, the estrous ovulation rates with Synchromate-B, Veramix and CIDR were 2.11 $\pm$ 1.89, 1.35 $\pm$ 0.87 and 1.43 $\pm$ 0.96, respectively. The estrous ovulation rates of the Synchromate-B treatment were significantly higher than those of the other treatments (P<0.05). 3. The average superovulation rates were 11.76 $\pm$ 6.00 and 11.07 $\pm$ 6.46 for the PMSG treatment and control groups, showing that there was no PMSG effects for the superovulation treated with CIDR.

• Journal of Embryo Transfer
• /
• v.16 no.2
• /
• pp.79-89
• /
• 2001

Steroid hormones control the expression of many cellular regulators, and a role thor estrogen in mouse oocytes has been well documented. The preovulatory $E_2$increment is generally accepted as the endocrine process regulating induction of in vivo oocyte maturation To address whether the activity of the T-type $Ca^{2+}$ channel is altered by 17 beta-estradiol ( $E_2$), we examined the actions of $E_2$on the calcium channel of mouse oocytes and early embryos. Oocrtes were collected from the oviduct of mice treated with pregnant mare's serum gonadotropin (PMSG) and human choronic gonadotropin (hCG). Whole cell voltage clamp technique and confocal microscopy were used to examine that $E_2$increase intracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{i}$ ) via voltage dependent $Ca^{2+}$ channel (VDC) and estrogen receptor (FSR), and $E_2$concentration by the use of radioimmunoassay (RIA) were examined in mouse. The results obtained were as follows: The peak of $Ca^{2+}$ current induced by $E_2$increased 122% to 1.50$\pm$0.03 nA from 1.23$\pm$0.21 nA (n=15) in the presence of 5 mM extracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{o}$ ). The increased $Ca^{2+}$ current was temporally associated with $Ca^{2+}$ transients. The intracellular $Ca^{2+}$ level increased 207%~30 s following the addition of 1${\mu}{\textrm}{m}$ $E_2$(relative fluorescence intensity: 836.4$\pm$131.2 for control, n=10, 1736.4$\pm$192.0 in the presence of $E_2$, n=10). $E_2$increased amplitude of $Ca^{2+}$ current and [C $a^{2+}$]$_{i}$ . $E_2$-induced $Ca^{2+}$ current and $E_2$concentration in blood were showed difference on the stage of embryo. These results suggest that $E_2$modulate $Ca^{2+}$ channel to increase $Ca^{2+}$ influx.$Ca^{2+}$ influx.

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2004.10a
• /
• pp.118-118
• /
• 2004

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2002.11a
• /
• pp.106-106
• /
• 2002

미토콘드리아는 세포내의 에너지대사에서 중요한 역할을 수행하는 세포내 소기관이며, 자체의 유전물질이 모계를 통해 유전되는 특징을 가지고 있다. 포유류 초기 배아의 발생과정에서 미토콘드리아의 역할과 기능에 대한 연구는 미진한 상태이다. 본 연구에서는 생쥐 초기 배아의 발생과정에서 관찰할 수 있는 미토콘드리아 미세구조의 변화 양상을 살펴보고, 이와 초기 배아의 발생 능력과의 관련성을 밝혀보고자 하였다. 과배란 유도된 ICR 생쥐로부터 배란된 난자와 2-세포기 배아를 수획하여 76 배양액으로 포배기까지 체외배양하면서, 각각의 발생단계에 따라 시료를 수획하였다. 미토콘드리아 미세구조의 변화는 일반적인 투과전자현미경방법(TEM)을 이용하여 관찰하였다. 미토콘드리아의 미세구조는 배란 난자에서 4-세포기 배아까지는 구형이고 크리스타가 발달하지 않은 원시형태였지만, 포배기로 발달함에 따라 크리스타가 발달된 막대형의 전형적인 미토콘드리아로 분화됨이 관찰되었다. 체외배양 중에 발생이 지연되거나 정지된 배아에서 관찰한 미토콘드리아의 미세구조는 공포화 (vacuolization), 크리스타 발달 지연, 손상된 미토콘드리아의 세포막 등과 같은 비정상적인 변형을 관찰할 수 있었다. 또한, 극체에 존재하는 미토콘드리아의 미세구조는 정상적인 핵내의 유전자와의 상호작용이 없어 미분화 상태로 포배기까지 유지되는 것을 관찰하였다. 이상의 결과를 통해 미토콘드리아의 정상적인 분화 과정이 초기 배아의 발생능력과 관련되어 있음을 알 수 있었으며, 포유류 초기 배아의 체외배양시스템을 개선하는데 미토콘드리아 미세구조의 관찰과 변화에 대한 고려가 있어야 될 것으로 생각된다. buffer A 용액으로 세척하여 유리 petri dish의 바닥에 부착된 macrophage만을 cell scraper로 분리하였다. 분리한 macrophage는 0.5-1 $\times$ $10^{6}$ cells/$m\ell$가 되게 조정하여, IL-I 을 0.001, 0.01, 0.1 또한 1 ng/$m\ell$를 첨가하여 농도에 따른 효과를 조사하였고, 각각 24, 48, 72, 96 또한 120시간을 배양하여 시간에 의한 효과도 실시하였다. 각 배치구에서 얻어진 배양액은 TGF-$\beta$를 조사하기 전까지 -2$0^{\circ}C$에 동결 보존하였다. TGF-$\beta$의 측정은 TGF-$\beta$ kit(promega, USA)를 이용하여 실시하였으며, 통계학적 분석은 Anova test를 Statview program을 이용하여 분석하였다. 시험의 결과 대조구에 비해 IL-I 첨가구는 2-3배의 TGF-$\beta$생산을 보였으며, 배양시간에 따른 생산은 시간이 지남에 따라 약간 상승하는 경향을 보였으나, 유의적인 차이를 보이지는 않았다. 또한 IL-I의 농도에 따른 생산의 변화는 IL-I의 농도에 따라 약간의 차이를 보였고 유의적인 차이는 인정되지 않았다. 임신 및 비임신의 경우 임신우의 비장 macrophage가 비임신보다는 약간 상승하는 거스로 나타났다. 이상의 결과로 볼 때 IL-I $\alpha$ 는$\bet

• Clinical and Experimental Reproductive Medicine
• /
• v.36 no.2
• /
• pp.137-142
• /
• 2009

Ovarian hyperstimulation syndrome (OHSS) is one of the most common iatrogenic complications induced by assisted reproductive technology. Hydrothorax develops in about 10 percent of patients with severe OHSS and it is usually associated with marked ascites. On the other hand, severe hydrothorax without marked ascites is uncommon, and its pathogenesis remains unclear. We recently experienced a case of severe unilateral hydrothorax with minimal ascites induced by quintuplet pregnancy following intrauterine insemination with controlled ovarian hyperstimulation. Severe hydrothorax was resolved after only conservative and symptomatic management without invasive procedure such as thoracentesis or paracentesis. We report this case with brief review of literature.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.88-88
• /
• 2003

본 연구는 배란후 48시간째 회수된 난소로부터 황체세포를 체외배양 후 progesterone 분비에 대한 LH, IGF-1 및 TGF$\beta$첨가효과에 대해 검토하였다. 축산기술연구소에서 사육중인 돼지(체중 145$\pm$kg) 12두로부터 발정을 유도시켜 배란후 약 48시간째 도축하여 난소를 회수하였다. 회수된 난소로부터 황체를 분리하여 세절한 후 0.25% collagenase용액(0.025mg DNase, 50mM EDTA, 50mM Dithiothreitol)으로 37$^{\circ}C$의 진탕수조에서 30분간 배양하여 황체세포를 분리 회수하였다. 회수된 황체세포는 D-MEM 용액(GIBCO, 10% FCS와 antibiotics 첨가)으로 2회 세정하여 1$\times$$10^{6}$live cell/$m\ell$이 되도록 희석한 후 24 well culture plate(Coming, New Tork 14831)에 분주하여 $CO_2$ 배양기($CO_2$: 5%)에서 12시간 간격으로 배양액을 회수하였으며 48시간까지 배양하였다. 배양액 1$m\ell$당 LH, IGF-I 및 TGF$\beta$1을 단독 혹은 공배양하여 회수된 배양액내의 progesterone 농도를 RIA법으로 분석하였다. 돼지황체세포는 배양후 24시간째에 높은 농도의 progesterone이 측정되었으며 LH를 첨가한 대조구에 비해 유의적으로 높은 progesterone이 측정되었다. 또한 IGF-1 과 TGF$\beta$1을 첨가한 군에서도 높은 농도의 progesterone이 측정되었다 그러나 TGF$\beta$ 혹은 IGF-I 단독으로는 대조구의 결과와 큰 차이가 없었다 따라서 돼지 황체세포에서의 progesterone 분비는 TGF$\beta$ 및 IGF-I 그리고 LH의 황체세포의 progesterone 분비기능을 촉진하는 것으로 나타났다.

• Clinical and Experimental Reproductive Medicine
• /
• v.15 no.1
• /
• pp.25-34
• /
• 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.