• 한국수정란이식학회지
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• 제19권2호
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• pp.113-119
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• 2004

본 연구는 재래산양에 있어서 과 배란 처리에 의한 oocyte의 회수방법과 양질의 oocyte 회수 체계를 확립하기 위하여 과 배란 처리 후 회수시간이 난포의 발달과 난자의 회수율에 미치는 영향을 조사하였다. 공시동물은 체중 15~25 kg 전${\cdot}$후의 성숙한 미경산 재래산양으로서 발정동기화를 위하여 CIDR를 10일간 질내에 삽입하고 과배란 처리는 FSH를 CIDR 삽입 8, 9, 10일째에 12시간 간격으로 70 mg을 감량 투여하였다. $PGF_{2\alpha}$는 FSH와 함께 8일째에 투여하였으며, CIDR는 10일째에 제거와 동시에 hCG 400 IU를 투여하였다. 난자의 회수는 hCG 투여 후 29~50시간째에 시간대별로 외과적인 방법으로 실시하였다. hCG 투여 후 회수시간에 따른 in vivo란의 회수에 있어서 각각 5.93${\pm}$2.88(29~34시간), 6.82${\pm}$0.95(35~40시간) 및 7.33${\pm}$1.54개(41~50시간)로서 차이가 없었다. 회수율에 있어서는 35~40시간째에 회수하였을 때가 49.7%로서 29~34시간 및 41~50시간째에 38.2% 및 29.5% 보다 유의적(P<0.05)으로 높았다. 두당 회수 난자 수는 각각 2.27${\pm}$0.76, 3.39${\pm}$0.75 및 2.17${\pm}$1.40개로서 차이가 없었다. hCG 투여 후 29~34, 35~40, 및 41~50시간에 관찰된 난포 수는 두당 11.75${\pm}$2.45, 11.87${\pm}$1.34 및 9.20${\pm}$1.50개였다. 또한 배란된 성숙난자의 채란 율은 각각 70.2, 74.7 및 54.3%로서 41~50시간째에 회수하였을 때가 가장 낮았다. 두당 회수율에 있어서도 8.25${\pm}$1.34, 8.87${\pm}$1.10 및 5.00${\pm}$1.30개로서 회수시간에 따른 유의적인 차이는 없었다. 회수한 난포내 미성숙 난자의 등급에 있어서 회수시간대별 1등급은 각각 24.2, 19.5 및 12.0%였으며, 2등급의 경우는 41~50시간이 4.0%로서 29~34시간과 35~40시간의 14.4% 및 16.2%보다 유의적(P<0.05)으로 낮았다.

• Clinical and Experimental Reproductive Medicine
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• 제26권2호
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• pp.163-170
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• 1999

Objectives: The aims of this study are 1) to determine if GAST is a better indicator in predicting ovarian response to COH compared with patient's age or basal FSH level and 2) to evaluate its role in detecting abnormal ovarian response. Design: Prospective study in 118 patients undergoing IVF-ET using GnRH-a short protocol during May-September 1995. Materials and Methods: After blood sampling for basal FSH and estradiol $(E_2)$ on cycle day two, 0.5ml (0.525mg) GnRH agonist ($Suprefact^{(r)}$, Hoechst) was injected subcutaneously. Serum $E_2$ was measured 24 hours later. Initial $E_2$ difference $({\Delta}E_2)$ was defined as the change in $E_2$ on day 3 over the baseline day 2 value. Sixteen patients with ovarian cyst or single ovary or incorrect blood collection time were excluded from the analysis. The patients were divided into three groups by ${\Delta}E_2$; group A (n=30):${\Delta}E_2$<40 pg/ml, group B (n=52): 40 pg/ml${\leq}{\Delta}E_2$<100 pg/ml, group C (n=20): ${\Delta}E_2{\leq}100$ pg/ml. COH was done by GnRH agonist/HMG/hCG and IVF-ET was followed. Ratio of $E_2$ on day of hCG injection over the number of ampules of gonadotropins used ($E_2hCGday$/Amp) was regarded as ovarian responsiveness. Poor ovarian response and overstimulation were defined as $E_2$ hCGday less than 600 pg/ml and greater than 5000 pg/ml, respectively. Results: Mean age $({\pm}SEM)$ in group A, B and C were $33.7{\pm}0.8^*,\;31.5{\pm}0.6\;and\;30.6{\pm}0.5^*$, respectively ($^*$: p<0.05). Mean basal FSH level of group $A(11.1{\pm}1.1mlU/ml)$ was significantly higher than those of $B(7.4{\pm}0.2mIU/ml)$ and C $(6.8{\pm}0.4mIU/ml)$ (p<0.001). Mean $E_2hCGday$ of group A was significantly lower than those of group B or C, i.e., $1402.1{\pm}187.7pg/ml,\;3153.2{\pm}240.0pg/ml,\;4078.8{\pm}306.4pg/ml$ respectively (p<0.0001). The number of ampules of gonadotropins used in group A was significantly greater than those in group B or C: $38.6{\pm}2.3,\;24.2{\pm}1.1\;and\;18.5{\pm}1.0$ (p<0.0001). The number of oocytes retrieved in group A was significantly smaller than those in group B or C: $6.4{\pm}1.1,\;15.5{\pm}1.1\;and\;18.6{\pm}1.6$, respectively (p<0.0001). By stepwise multiple regression, only ${\Delta}E_2$ showed a significant correlation (r=0.68, p<0.0001) with $E_2HCGday$/Amp, while age or basal FSH level were not significant. Likewise, only ${\Delta}E_2$ correlated significantly with the number of oocytes retrieved (r=0.57, p<0.001). All four patients whose COH was canceled due to poor ovarian response belonged to group A only (Fisher's exact test, p<0.01). Whereas none of 30 patients in group A (0%) had overstimulation, 14 patients among 72 patients (19.4%) in group B and C had overstimulation (Fisher's exact test, p<0.01). Conclusions: These data suggest that initial $E_2$ difference after GAST may be a better prognostic indicator of ovarian response to COH than age or basal FSH level. Since initial $E_2$ difference demonstrates significant association with abnormal ovarian response such as poor ovarian response necessitating cycle cancellation or overstimulation, GAST may be helpful in monitoring and consultation of patients during COH in IVF-ET cycle.

• 대한한방부인과학회지
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• 제25권2호
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• pp.43-65
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• 2012

Objectives: This study was designed to investigate the effects of Hyeolbuchukeotang( HCT) on the Estradiol Valerate(EV) induced Polycystic Ovaries and the Implantation of rats. Methods: PCO was induced by single intramuscular(IM) injection with EV in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for 60 days. Control group(n=8) were injected with EV and orally administrated distilled water for 60 days. HCT treated group(n=8) were injected with EV and orally administrated HCT for 60 days. At the end day of experiment, we measured weights of body, ovaries, adrenal glands, uterus and contents of serum LH, FSH, ADD. The histomorphometrical changes of ovaries were also evaluated. And we observed the NGF and CRF expression by immunohistochemistry. Also we observed the mating and count the number of implantation. Results: - The weights(mg) of ovaries in HCT treated group($58.9{\pm}6.5$) were significantly increased (p<0.01) compared with control group ($42.3{\pm}8.5$). - The number of mature follicles in HCT treated group($9.8{\pm}2.6$) was significantly increased (p<0.01) compared with control group($6.1{\pm}2.1$). - The number of cystic follicles in HCT treated group($1.6{\pm}1.2$) was significantly decreased (p<0.01) compared with control group($3.8{\pm}1.5$). - The value of serum FSH(mIU/ml) in HCT treated group($3.870{\pm}2.151$) was significantly decreased(p<0.05) compared with control group($10.476{\pm}6.294$). - The expression of NGF-immunoreactive cells in the ovarian cells in HCT treated group was lesser observed than control group. - The number of implantation in HCT treated group($7.4{\pm}1.8$) was significantly increased (p<0.05) compared with control group($3.1{\pm}4.4$). Conclusions: Hyeolbuchukeo-tang(HCT) is effect on polycystic ovaries and the implantation of rats by EV-induced.

• 한국수정란이식학회지
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• 제14권2호
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• pp.121-129
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• 1999

The objective of this study was to produce calves by transfer of embryos derived from slaughter house(SHD) and ultrasound-guided ovum pick-up (OPU). At 60 hrs after injection of 400 mg FSH dissolved in 25% polyvinylpyrrolidone(PVP) by single dose, ultrasound-guided follicular oocyte aspiration was ferformed. Day-7 and day-8 blastocysts produced by in vitro maturation (IVM), fertilization (IVF) and culture(IVC) of the oocytes derived from SHD and OPU were nonsurgically transferred into recipients. The results obtained were as follows. The cleavage rate and the development rate to blastocysts were not significantly (P<0.05) different between the oocytes obtained by SHD (72.9% vs. 34.1%) and OPU (75.9% vs. 38.4%). The oocyte recovery rate from the number of follicles by ultrasound-guided aspiration were not significantly (P<0.05) different between Holstein (61.7%) and Hanwoo (60.1%), but the rate of oocytes useful for IVF was significantly (P<0.05) higher in Hanwoo (69.3%) than Holstein (59.6%). The cleavage rate and the development rate to blastocysts was not significantly (P<0.05) different between Holstein (74.9% vs. 39.2%) and recipients on day 8 of estrus cycle resulted in 13 pregnancies (34.2%). One of them was sacrificed during gestation period due to mastitis and another was aborted spontaneous. The resulting 14 calves were morphologically normal at birth. Seventy eight fresh OPU-IVF embryos were transferred into 21 recipients on day 8 of estrus cycles, resulting in pregnancy of 12 recipients (41.4%). Two of them were sacrificed during gestation period due to mastitis and the other two were aborted. Nevertheless, the 11 OPU-calves have been born normally.

• Reproductive and Developmental Biology
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• 제28권3호
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• pp.181-185
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• 2004

본 연구는 동물복제 및 형질전환 동물생산 등의 연구에 기초자료를 제공하고자 재래산양의 oocyte를 단위발생을 유도하여 회수난자의 조건, 활성화방법, 배양조건 등이 단위발생란의 체외발달율에 미치는 영향을 조사하여 재래산 양의 최적의 배양조건을 확립하고자 실시하였다. 난자의 회수는 체중 15～25 Kg 전 ㆍ 후의 성숙한 미경산 재래산양에 FSH와 PMSG 를 사용하여 과배란을 유기하여 hCG 투여 후 제 35시간째에 외과적인 방법으로 in vivo (체내성숙) 난자는 난관을 관류하는 방법으로 회수하였고, in vitro (체외성숙) 난자는 난포로부터 흡입하여 난포란을 채취하여 약 22시간 체외성숙을 실시하였다. 활성화 처리는 전기자극법과 Ionomycin + 6-DMAP를 처리하여 단위발생을 유도하였다. 복제수정란의 배양은 M16, TCM-199 및 mSOF 배양액으로 6～7일 동안 체외배양을 실시하였다. 활성화를 유도하기 위하여 전기자극 및 ionomycin + 6-DMAP 처리를 하였을 때 분할율은 각각 64.1 및 76.5%로서 이들간에 차이는 없었다. 배반포기로의 발달율은 전기자극방법으로는 전혀 발달이 이루어지지 않았으나, ionomycin +6-DMAP 처리방법에서는 15.6%가 배반포로 발달하였다. In vivo 난자와 in vitro 난자를 활성화를 유도하였을 때 분할율은 86.8 및 69.0%로서 이들간에 유의적인 차이는 없었다. 4-세포기(93.9% vs 66.1%), 8-세포기(90.9% vs 37.0%) 및 상실배기(89.4% vs 23.6%)는 이들간에 유의적(P<0.05)인 차이가 있었으며, 배반포기로의 발달율은 체내성숙난자가 50.0%로서 체외성숙난자의 0.8%보다는 유의적으로 높았다. 활성화를 유도한 난자를 mSOF 배양액으로 체외배양을 실시하였을 때 분할율은 81.0%로서 TCM-199 +oviduct cell 의 64.3% 및 Ml6 배양액의 51.6%보다는 높게 나타났다. 배반포기로의 발달율은 mSOF 배양액에서는 3.4%가 발달하였으나, TCM-199+ oviduct cell 배양액과 M16 배양액에서는 전혀 발달이 이루어지지 않았다. 이상의 결과는 포유동물 난자의 단위발생의 유도 및 체외배양시 난자의 공급원, 난자의 활성화 방법 및 배양조건 등이 차후 단위발생란의 체외발생율에 크게 영향을 미칠 수 있는 근거를 제시해 준다.