• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.137-148
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• 1999

Purpose of the present study was to find the optimal ovulation induction medicine for the maturation and development of immature oocytes and culture media for 2-cell embryos in the mouse model. ICR female mouse aged 6 to 8 weeks, were stimulated with 5 IU PMSG injection. At 47 to 50 hour post-PMSG injection, ovaries were dissected out and oocytes-cumulus complexes were punctured. The oocyte-cumulus complexes were cultured in media containing various ovulation induction medicine, CC, HMG and Metrodin for 18 hours. Female ICR mice were stimulated with 5 IU PMSG and 48 hours later were injected 5 IU of hCG, then female and male mice were mated. At 48 hour post-hCG injection, oviducts were dissected out and 2-cell embryos were flushed. The 2-cell embryos were cultured in various media, Ham's F-10 media of milli-Q water $(3^{\circ})$, Ham's F-10 media of HPLC (high performance liquid chromatography, Baxter) water, Medicult media, HTF (human tubal fluid) media for 96 hours. The results were as follows. 1. When the oocytes-cumulus complexes were cultured in $10^{-9}{\mu}g/ml{\sim}10^{-8}{\mu}g/ml$ of CC, those were suppressed in meiotic maturation $(28.2{\sim}33.7%)$. Whereas the oocytes-cumulus complexes were cultured in $10^{-7}{\mu}g/ml{\sim}10^{-4}{\mu}g/ml$, these were not effected in meiotic maturation $(54.5{\sim}72.7%)$. 2. When the oocytes-cumulus complexes were cultured in $10^{-4}{\mu}g/ml{\sim}10^{-1}{\mu}g/ml$ of Metrodin, those were suppressed in meiotic maturation $(35.7{\sim}41.5%)$. Meanwhile the oocytes-cumulus complexes were cultured in $10^{-7}{\mu}g/ml{\sim}10^{-5}{\mu}g/ml$, those were not effected in meiotic maturation $(54.2{\sim}70.3%)$. 3. When the oocytes-cumulus complexes were cultured in $10^{-5}{\mu}g/ml{\sim}10^{-4}{\mu}g/ml$ of HMG, those were suppressed in meiotic maturation $(48.2{\sim}50.4%)$. As being cultured in $10^{-7}{\mu}g/ml{\sim}10^{-6}{\mu}g/ml$, increased in meiotic maturation $(75.8{\sim}80.7%)$. 4. When the 2-cell embryos were cultured in Ham's F-10 media of milli-Q water $(3^{\circ})$, Ham's F-10 media of HPLC (high performance liquid chromatograpy, Baxter) water, Medicult media, HTF (human tubal fluid) media, developmental rates to blastocyst and hatching for 96 hour were 50.0%, 45.2%, 71.5% and 95.6%, respectively.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.253-258
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• 1997

토끼에서는 수정란 이식과 같은 기본적인 번식공학적 방법의 효율성이 아직 생쥐와 같은 실험동물에 비해 떨어지고 있어 생물공학적인 기술을 응용하는데 큰 어려움이 있다. 특히 유전자 이식에 의한 형질전환 토끼의 생산과 같은 생물공학적인 기술을 실용화하는데 효율이 높은 수정란이식 기술의 개발이 필수적이라고 할 수 있다. 본 연구에서는 토끼에서 수정란 이식 기술의 첫 단계인 과배란 유도를 효율적으로 이용할 수 있는 방법을 정립하기 위해 반복적인 과배란 유도가 배란율 및 수정란의 질적인 면과 양적인 면에 미치는 영향을 조사하였다. 연구방법으로는 FSH와 HCG를 사용하여 과배란을 유도하였고 2.5 개월의 반복처리간격으로 3번의 반복적인 과배란 처리를 한 후 반복처리에 따른 배란율과 배란된 난자의 형태학적 상태, 배양에 의한 발생 능력 상태 등을 감소하였으며 배란수의 변이도 커지는 경향을 나타내었다(첫번째, 32.6(+-)2.5; 두번째 28.7(+-)3.7; 세번째 20.99(+-)3.8). 제 2극체의 돌출, 전핵의 형성, cummulus cell의 존재등에 의한 회수된 난자의 형태학적 관찰에 의한 방법으로 난자를 분류한 결과 과배란의 반복수가 증가함에 따라 다양한 모양의 난자가 회수되어 배란이 광범위한 시간대에 일어나고 있음을 나타내었다. 또한 과배란의 반복적인 유도에 의해 난소의 혈포수는 증가하였으나 채란된 난자의 채외배양에 의한 발육율에는 차이가 없었다. 그러므로 과배란의 반복적인 유도는 공란토의 난소반응에는 영향을 미쳤으나 난자의 질에는 영향을 미치지 않았음을 나타낸다.

• Journal of Aquaculture
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• v.10 no.1
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• pp.55-61
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• 1997

The experiment was conducted to understand the effect of HCG upon ovulation induction and egg maturation in the sevenband grouper, Epinephelus septemfasciatus, as a part of the research for its resources management and seed production. Three different size groups of sevenband groupers showing 300~400g, 1,500~2,000g and 2,500~4,000g, were collected from June to August, 1996. They were treated two or three times with HCG (500~1,000 IU/kg). Gonadal development of the sevenband groupers were examined using the paraffin section method and then stained with haematoxylin and eosin. When smaller groups of sevenband grouper having 300~400g, 1,500~2,00g and 2,500~3,200g in weight were treated two or three time with HCG 9500~1,000 IU/g), their gonads contained oogonia, oocytes with the size of 20~30${\mu}{\textrm}{m}$, and stroma tissues. However, in case of two HCG treatments with a dosage of 1,000 IU/kg on sevenband groupers of 3,400~4,000g, both healthy eggs of $800\mu$m in diameter and regressing eggs were ovulated. A sevenband grouper of 4,000g obvulated 4,252 eggs in total. Of these, healthy and regressing eggs turned out to be 20.8% and 79.2%, respectively. In addition, previtellogenic oocytes of around $50\mu$m, oocytes of 20~$30\mu$m, and ovulation trace were also observed in the gonad.

• Korean Journal of Animal Reproduction
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• v.8 no.1
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• pp.36-45
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• 1984

This study was carried out to investigate the changes in ovary in repeatedly superovulated rabbits. A total of 57 New Zealand White and Californian, 25 mature virgin and 32 immature does were used in this study. For induction of repeated superovulation, PMSG and HCG were injected at 17-day and 30-day intervals for mature does and 17-day intervals for immature ones. The repeatedly superovulated does at 17-day intervals were induced luteolysis of pseudopregnant corpus luteum with PGF2${\alpha}$ on Day 8 to 9 p.c. The effect of repeated superovulation on reproductive organs was investigated on Day 3 p.c. in mature does and on Day 3 and 6 p.c. in immature ones, respectively. 1. In mature virgin does, the number of ovulation points in the 2nd and 3rd superovulation period averaged 7.0 and 5.0 at 17-day intervals and 13.4 and 6.0 at 30-day intervals, respectively. These numbers were statistically similar to 9.5 ovulation points in the control. However, there were less (p<0.05) ovulation points in those periods compared with 22.1 ovulation points in the 1st superovulation period. 2. In immature does, the number of ovulation points in the 2nd and 3rd superovulation period averaged 5.3 and 2.3, respectively. These numbers were significantly (p<0.05) decreased than 17.1 ovulation points in the 1st periods. The number of ovulation points in the 2nd superovulation period was similar to that in the control, but there was a significant (p<0.05) decrease in the number of ovulation points in the 3rd period as compared to the control. 3. In mature virgin does, the number of visible normal and hemorrhagic follicles (>1.0mm diameter) on day 3 p.c. averaged 19.1 and 8.9 in the 1st superovulation period, respectively. In the 2nd 3rd superovulation period, the number of normal follicles averaged 8.3 and 15.5 at 17-day intervals and 17.8 and 14.5 at 30-day intervals. The number of hemorrhagic follicles in the 2nd and 3rd superovulation period averaged 6.3 and 2.0 at 17-day intervals and 5.2 and 7.8 at 30-day intervals, respectively. There was a slight decrease, although not significant, in the number of normal and hemorrhagic follicles in the 2nd and 3rd period at 17-day intervals compared to that in the 1st period. 4. In immature does, the number of visible normal follicles on day 3 and day 6 p.c. in the 1st superovulation period averaged 27:3 and 26.1, respectively. The follicles on day 3 p.c. tended to increase slightly more than that in the cortrol, but the average number of normal follicles on day 6 p.c. did not differ from that in the control. The number of visible hemorrhagic follicles on day 3 and day 6 p.c. in the 1st of follicles in the 1st superovulation period average 10.2 and 9.9, respectively. There was a slight increase in the number of follicles in the 1st period compared to that in the control. In the 2nd and 3rd superovulation period, the number of normal follicles revealed a slight decrease in the 3rd period, but the number of hemorrhagic follicles was not different between periods. 5. The number of growing follicles with incipient intral formation on day 3 p.c. in mature does of the 1st superovulaton period average 29.7 and the average number of growing follicles in the 3rd period was 26.7 at 17-day intervals and 31.0 at 30-day intervals, respectively. These numbers did not differ from that in the control. In immature does, the number of growing follicles averaged 57.7, 45.0 and 59.3 in the 1st, 2nd and 3rd superovulation period, respectively. There was a slight but not significant decrease in the number of growing follicles in the 3rd period compared to that in the control.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.1
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• pp.27-32
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• 2005

Objective: To evaluate the effectiveness of aromatase inhibitor (AI) for ovulation induction in polycystic ovary syndrome (PCOS) patients with thin endometrium, hyper-responsiveness after clomiphene citrate (CC) treatment. Material and Methods: A prospective study was performed in 43 PCOS patients (50 cycles) with ovulatory dysfunction between March 2004 and September 2004. AI group (total 36 cycles) included the patients 1) with thin endometrium below 6 mm on hCG day after CC (n=17), 2) with more than 5 ovulatory follicles after 50mg of CC (n=4), 3) who do not want multiple pregnancy (n=14). Patients were treated with Letrozole 2.5mg for days 3 to 7 of the menstrual cycle. CC group (total 14 cycles) were treated with CC 50~100 mg. Results: In PCOS patients, ovulation was occurred 97.2% after AI use. Between AI group and CC group, there was no significant difference in the mean age, duration of infertility, interval of menstruation, basal FSH, prior treatment cycles, and the day of hCG administration. But, the number of mature follicles (${\geq}15mm$) was lower in the AI group ($1.08{\pm}0.45$ vs. $1.64{\pm}0.75$) (p=0.018), and the thickness of endometrium (mm) was significantly thicker in the AI group ($10.35{\pm}1.74$ vs. $9.23{\pm}1.61$) (p=0.044), and E2 (pg/ml) concentration on hCG day was lower in the AI group ($116.9{\pm}75.8$ vs. $479.5{\pm}300.8$) (p=0.001). Among the AI group, patients with prior thin endometrium (below 6 mm) during CC treatment showed $10.6{\pm}1.6mm$ in the endometrial thickness and $106.6{\pm}66.8pg/ml$ in $E_2$ concentration. Patients with more than 5 ovulatory follicles after CC showed decreased follicle number ($1.25{\pm}0.5$) compared to prior CC cycle. Conclusions: In PCOS patients, AI group showed significantly thicker endometrium, lesser number of mature follicles, and lower E2 concentration on hCG day than CC group. AI might be useful alternative treatment for ovulation induction in PCOS patients with thin endometrium and hyper-responsiveness after CC treatment.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.169-176
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• 2009

Domestic bitches are non-seasonally monoestrus; spontaneously ovulate only once or twice occurs at anytime of the year. Estrus induction has been applied infrequent estrus, misleading ovulation, mating difficulties, failure to conceive after normal mating, pregnancy failure and biological research. Protocol of estrus induction which included variable hormones such as FSH, GnRH, and PMSG have been applied for the last decades. Recently, Bromocriptine, one of anti-prolatin/dopamine agonist has been occasionally applied for estrus induction. The study was carried out to investigate the effective method for the induction of estrus in bitches using different hormone treatments, and the initiation time of estrus from hormone treatment by assessments of cytological observation and blood plasma progesterone concentration. A total of 54 bitches on anestrus were selected for the study and divided randomly into 8 treatment groups as follow. Control, natural estrus; FSH (L), FSH (1.5 mg/kg, twice a day, $Falltrophin^{(R)}$, Vetrepharm); FSH (H), FSH (3.0 mg/kg, twice a day); GnRH+FSH, GnRH (5 ug/kg, once first day, $GNADON^{(R)}$, Dongbang)+FSH (3 mg/kg, SID); PMSG, PMSG (50 IU/kg, every third day $FOLLIGON^{(R)}$, Intevet); GnRH+PMSG, GnRH (5 ug/kg, only first day)+PMSG (50 IU/kg, every third day); GnRH, GnRH (5 ug/kg, only first day); Bromocriptine, bromocriptine (0.3 mg/kg, SID, $Parlodel^{(R)}$, Novartis). The bitches were evaluated clinical sign, cytological exam and $OVUCHECK^{(R)}$ Premate for assessment of estrus induction. Estrus induction rates were significantly (P<0.05) higher in GnRH+PMSG (100%) compared to others. PMSG and GnRH+PMGS (87.5 and 100%) and Bromocriptine (77.8%) were higher than others except GnRH+PMSG. Analysis of vaginal smear has proved to be effective a correct assessment of estrus induction with assay of progesterone concentration by $OVUCHECK^{(R)}$ Premate. Proestrus initiated by the $6^{th}$ after induction in most case. In conclusion, bromocriptine is an effective drug for estrus induction in bitches and assay of progesterone concentration by $OVUCHECK^{(R)}$ Premate with examination of vaginal smear that should be useful to detection of estrus induction of estrus induced bitches.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.1
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• pp.54-57
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• 2016

Controlled ovarian hyperstimulation is one of the major steps of in vitro fertilization. The inaccessibility or non-visualization of developing follicles on transvaginal sonography (the preferred imaging method) may be misjudged as a poor response, resulting in cycle cancellation. It is necessary to scrupulously appraise proxy indicators for ovarian response, such as estradiol levels, endometrial thickness, and other individual clinical characteristics. This can prompt meticulous transabdominal ultrasound follicular monitoring and oocyte retrieval with the goal of averting cycle cancellation and improving treatment outcomes.

• The Korean Journal of Cytopathology
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• v.7 no.1
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• pp.51-58
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• 1996

The cytologic samples of 26 ovarian cystic lesions from 25 women, aspirated under guide by trans-vaginal ultrasound, were evaluated for clinicopathologic correlation. Clinically 20 women were seeking medical assistance for infertility problems, and trans-vaginal cyst aspiration was done during follow-up of ovulation induction. Among them seven cases were histologically confirmed. Twenty cases of "benign cyst" in cytologic diagnosis were follicullar cyst and two cases of "endometriotic cyst" in cytology were histologically also proven in one case. One false positive diagnosis was given to corpus luteum cyst. It is emphasized that because the cysts are aspirated transvaginally and mature squamous epithelial cells occasionally could be mixed in the sample, attention should be given not to diagnose such cases as mature cystic teratoma. According to this study. ovarian aspiration cytology is useful in the management of cystic ovarian lesions, particularly in young women.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.4
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• pp.353-359
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• 2005

Heterotopic pregnancy is named when an extrauterine (ectopic) pregnancy coexists with an intrauterine pregnancy simultaneously by many causes such as PID (pelvic inflammatory disease), endometriosis, IUD (intrauterine device), previous pelvic surgery and others. This is very rare in general population, with a range of occurrence estimated between 1:7963 and 1:30000. But recently the incidence has increased as the uses of ARTs (assisted reproductive technologies) including ovulation induction, IVF-ET (in-vitro fertilization and embryo transfer) and GIFT (gamete intrafallopian transfer) increase. Because this has high maternal morbidity, mortality and fetal loss, early diagnosis and proper management is very important. We report a case of heterotopic pregnancy following IVF-ET with a brief review.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.207-212
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• 2020

Objective: Glutamate ionotropic receptor AMPA type subunit 1 (GRIA1) is a subunit of a ligand-gated ion channel that regulates the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by controlling the release of gonadotropin-releasing hormone. Few studies have investigated the association between the GRIA1 gene and human infertility. This study evaluated the association of the GRIA1 rs548294 C > T and rs2195450 G > A polymorphisms with the ovarian response to human menopausal gonadotropin (HMG) in Iranian women. Methods: One hundred women with histories of at least 1 year of infertility were included. On the second day of menstruation, patients were injected with HMG; on the third day, blood samples were collected. After hormonal analysis, the GRIA1 rs548294 C > T and rs2195450 G > A genotypes of samples were identified via the restriction fragment length polymorphism method, and on day 9, the number of follicles was assessed via ultrasound. Results: For the GRIA1 rs548294 C > T and rs2195450 G > A single nucleotide polymorphisms, the subjects with CT and GG genotypes, respectively, displayed the highest mean FSH level, LH level, and number of follicles on day 9 of the menstrual cycle (p< 0.05). Significant positive correlations were observed between LH and FSH (p< 0.01), LH and follicle count (p< 0.01), FSH and age (p< 0.05), follicle count and age (p= 0.048), and FSH and follicle count (p< 0.01). Conclusion: This study showed a significant relationship between GRIA1 polymorphisms and ovarian response to the induction of ovulation. Therefore, determining patients' GRIA1 genotype may be useful for improving treatment and prescribing suitable doses of ovulation-stimulating drugs.