• Journal of Animal Reproduction and Biotechnology
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• v.34 no.1
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• pp.35-39
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• 2019

This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.2
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• pp.62-66
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• 2015

Objective: To evaluate the effect of a gonadotropin-releasing hormone (GnRH) antagonist protocol using corifollitropin alfa in women undergoing assisted reproduction. Methods: Six hundred and eighty-six in vitro fertilization-embryo transfer (IVF)/ intracytoplasmic sperm injection (ICSI) cycles were analyzed. In 113 cycles, folliculogenesis was induced with corifollitropin alfa and recombinant follicle stimulating hormone (rFSH), and premature luteinizing hormone (LH) surges were prevented with a GnRH antagonist. In the control group (573 cycles), premature LH surges were prevented with GnRH agonist injection from the midluteal phase of the preceding cycle, and ovarian stimulation was started with rFSH. The treatment duration, quality of oocytes and embryos, number of embryo transfer (ET) cancelled cycles, risk of ovarian hyperstimulation syndrome (OHSS), and the chemical pregnancy rate were evaluated in the two ovarian stimulation protocols. Results: There were no significant differences in age and infertility factors between treatment groups. The treatment duration was shorter in the corifollitropin alfa group than in the control group. Although not statistically significant, the mean numbers of matured (86.8% vs. 85.1%) and fertilized oocytes (84.2% vs. 83.1%), good embryos (62.4% vs. 60.3%), and chemical pregnancy rates (47.2% vs. 46.8%) were slightly higher in the corifollitropin alfa group than in the control group. In contrast, rates of ET cancelled cycles and the OHSS risk were slightly lower in the corifollitropin alfa group (6.2% and 2.7%) than in the control group (8.2% and 3.5%), although these differences were also not statistically significant. Conclusion: Although no significant differences were observed, the use of corifollitropin alfa seems to offer some advantages to patients because of its short treatment duration, safety, lower ET cancellation rate and reduced risk of OHSS.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.3
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• pp.253-259
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• 2010

Objective: To evaluate if there is any correlation between the growth rate of dominant follicles and clinical characteristics or outcome variables in women undergoing controlled ovarian hyperstimulation (COH). Methods: This study was performed in 313 in vitro fertilization (IVF) cycles. Follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) were measured on day 3 of menstrual cycle, and serial ultrasonographic measurement of the diameter of growing follicles was performed. The growth rates of dominant follicles calculated by diameter difference divided by days were correlated with clinical characteristics and outcome variables. Results: There was no significant difference in the growth rate of the dominant follicles between gonadotropin releasing hormone (GnRH) agonist and antagonist cycles. No significant correlation was found between the growth rates and evaluated factors such as age, body mass index, LH, FSH, $E_2$, retrieved oocytes and fertilization rate. Conclusion: The Growth rate of dominant follicles seems to show an independent feature of basal characteristics and ovarian response.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.273-281
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• 1996

Previously, we have proposed a two-cell type model for follicular steroidogenesis inamphibians with Rana nigromacu lota. Present experiments were carried out to ascertain whether the model Is applicable to R. dybowskii. The role of theca layer were also reassessed by using granulosa cell-free pure theca layer (P-THEP). Theca/epithelium (THEP) layers, P-THEP layers, and granulosa cell enclosed-oocytes () were obtained from ovarian follicles of R. dybowskii by microdissection. Intact follicles (IFs) and different types of tissues were cultured for 6 hour in amphibian Ringer's m the presence or absence of FPII (0.05 gland/mi) or various steroid precursor (100 ng/ml). The amounts of product steroids converted by the components were measured by RIA. Exogenously added pregnenolone (P5) resulted in a marked increase in progesterone (P$_4$) by GCEOs (2143 pg/follicle) and IFs (2346 pg/follicle) but a smaller increase in P4 by THEP layer (495 pg/follicle). Addition of P$_4$ increased 17 a-hydroxyprogesterone (17 $\alpha$-OHP$_4$) levels by GCEOs (1118 pg/follicle) and IFs (1333 pg/follicle) but less by THEP layer (290 pg/follicle). However, much less amounts of P$_4$ or 17 $\alpha$-OHP$_4$ were producad by P-THEP layers than THEP in the presence of P5. Exogenous 1 7$\alpha$-OIIP$_4$ increased androstenedione (AD) levels by GCEOs (1415 pg/follicle) and IFs (561 pg/follicle) but not by THEP layers. In contrast, addition of AD resulted m a marked increase in testosterone (T) levels by TIIEP (2594 pg/follicle) and IFs (2223 pg/follide) but much less by GCEOs (339 pg/follicle). Exogenous T increased estradiol (E$_2$) levels by GCEOs (551pg/follicle) and IFs (887 pg/follicle), but not by THEP layer (<10 pg/follicle). Without addition of FPH or steroid precursors, very low or nondetectable levels of steroids were produced (< 20 pg/follicle) by all the types of follicular components examined. The data presented here indicate that the two-cell type model based on the study with R. nigromacu Iota is applicable to R. dybowskii and also suggest that the minor pathway, which convert P5 to 17$\alpha$-OHP$_4$, is not present in theca layer.

• Korean Journal of Animal Reproduction
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• v.13 no.2
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• pp.79-84
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• 1989

This study was carried out to investigate the effect of LH-RH and Gn-RH treatment in Holstein cows with ovarian quiescence and follicular cystic ovaries. The cows with ovarian quiescence and follicular cystic ovaries injected intramuscularly with 100$\mu\textrm{g}$, 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of LH-RH and 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of Gn-RH respectively. The cows was diagnosed by repeated rectal palpation. The plasma progesterone and estradiol-17$\beta$ concentrations were assayed by radioimmunoassay methods. The resutls of this study were summarized as follows : 1. Ovulations were induced after treatment of LH-RH and Gn-RH. The concentrations of progesterone reached small peak level at luteal phase and estradiol-17$\beta$ reached obvious peak level with the development and maturation of the follicle during the periods of degeneration of the corpus luteum, and normal ovarian cycle activity started subsequently. 2. The cows with ovarian quiescence and follicular cystic ovaries were induced ovulation at 38.9$\pm$5.3 hrs. after treatment of LH-RH in 66.7% cows and at 52.7$\pm$7.9 hrs after treatment of Gn-RH in 60.0% cows respectively. 3. The good ovarian responses were indicated in treatment with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of LH-RH than those treated with 100$\mu\textrm{g}$ in cows with ovarian quiescence, and did not show difference of ovarian responses between treatments with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of Gn-RH in cows with follicular cystic ovaries.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.241-245
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• 1994

Despite increasing success rate of IVF, poor response to ovarian stimulation remains a problem. So, attempts to improve ovarian responses, for example, by using combined gonadotropin-releasing hormone analogue(GnRH-a) and human menopausal gonadotropin(hMG) have shown limited success. It is reported that response of granulosa cells in vitro to FSH is stimulated by co-incubation with IGF-l, and IGF-l production can be increased by growth hormone. This suggest that combination regimen of G.H. and hMG may augment follicle recruitment. In fifteen patients who had previous history of poor ovarian response to gonadotropin stimulation after pituitary suppression with mid -luteal GnRH-a, the effectiveness of cotreatment with G.H. in IVF program was evaluated using a combination regimen of G.R. and hMG at Korea University Hospital IVF Clinic. Ovarian responses to gonadotropin stimulation in control and GH-treated cycles assessed by total dose and duration of hMG treatment, follicular development and peak $E_2$ level, number of eggs retrieved, and fertilization rates were also assessed. In each group, serum and follicular fluid IGF-1 concentrations on day of egg collection were measured by RIA after acidification and extraction by reveresed phase chromatography. Patients receiving G.H. required fewer days and ampules of gonadotropins, developed more oocytes, and more embryos transferred. But, the differences were not statistically significant, except the duration of hMG treatment. Our data showed a significantly higher concentration of IGF-l in the serum, not in the follicular fluid, of patients treated with G.H. compared with control group. These data suggest that growth hormone treatment does not improve the ovarian response in women with limited ovarian reserve to gonadotropin stimulation for IVF.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.1
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• pp.18-23
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• 2011

Objective: Peroxiredoxins (Prxs) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. This report examined the expression of Prx isotype I in the rat ovary after hormone treatment. Methods: Immature rats were injected with 10 IU of pregnant mare's serum gonadotropin (PMSG) to induce the growth of multiple preovulatory follicles and 10 IU of human chorionic gonadotropin (hCG) to induce ovulation. Immature rats were also treated with diethylstilbestrol (DES), an estrogen analogue, to induce the growth of multiple immature follicles. Northern blot analysis was performed to detect gene expression. Cell-type specific localization of Prx I mRNA were detected by in situ hybridization analysis. Results: During follicle development, ovarian Prx I gene expression was detected in 3-day-old rats and had increased in 21-day-old rats. The levels of Prx I mRNA slightly declined one to two days following treatment with DES. A gradual increase in Prx I gene expression was observed in ovaries obtained from PMSG-treated immature rats. Furthermore, hCG treatment of PMSG-primed rats resulted in a gradual stimulation of Prx I mRNA levels by 24 hours (2.1-fold increase) following treatment, which remained high until 72 hours following treatment. In situ hybridization analysis revealed the expression of the Prx I gene in the granulosa cells of PMSG-primed ovaries and in the corpora lutea of ovaries stimulated with hCG for 72 hours. Conclusion: These results demonstrate the gonadotropin and granulosa cell-specific stimulation of Prx I gene expression, suggesting its role as a local regulator of follicle development.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.2
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• pp.135-141
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• 2022

Objective: This study investigated the impact of two stimulation protocols using highly purified human menopausal gonadotropin (HP-hMG) on the endocrine profile, follicular fluid soluble Fas levels, and outcomes of intracytoplasmic sperm injection (ICSI) cycles. Methods: This prospective clinical trial included 100 normal-responder women undergoing ovarian stimulation for ICSI; 55 patients received concomitant follicle-stimulating hormone (FSH) plus HP-hMG from the start of stimulation, while 45 patients received FSH followed by HP-hMG during mid/late follicular stimulation. The primary outcome was the number of top-quality embryos. The secondary outcomes were the number and percentage of metaphase II (MII) oocytes and the clinical pregnancy rate. Results: The number of MII oocytes was significantly higher in the concomitant protocol (median, 13.0; interquartile range [IQR], 8.5-18.0 vs. 9.0 [8.0-13.0] in the consecutive protocol; p=0.009); however, the percentage of MII oocytes and the fertilization rate were significantly higher in the consecutive protocol (median, 90.91; IQR, 80.0-100.0 vs. 83.33 [75.0-93.8]; p=0.034 and median, 86.67; IQR, 76.9-100.0 vs. 77.78 [66.7-89.9]; p=0.028, respectively). No significant between-group differences were found in top-quality embryos (p=0.693) or the clinical pregnancy rate (65.9% vs. 61.8% in the consecutive vs. concomitant protocol, respectively). The median follicular fluid soluble Fas antigen level was significantly higher in the concomitant protocol (9,731.0 pg/mL; IQR, 6,004.5-10,807.6 vs. 6,350.2 pg/mL; IQR, 4,382.4-9,418.4; p=0.021). Conclusion: Personalized controlled ovarian stimulation using HP-hMG during the late follicular phase led to a significantly lower response, but did not affect the quality of ICSI.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.3
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• pp.200-205
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• 2023

Objective: Polycystic ovary (PCO), a diagnostic component of polycystic ovary syndrome (PCOS), requires either an ovarian volume (OV) criterion or a follicle number per ovary (FNPO) criterion. This study investigated the association of OV and FNPO criteria with various manifestations of PCOS. Methods: This cross-sectional study was conducted at a university hospital among 100 patients newly diagnosed with PCOS (according to the revised Rotterdam criteria). Fasting blood samples were collected to measure glucose, total testosterone (TT), luteinizing hormone (LH), follicle-stimulating hormone (FSH), lipid, insulin, and hemoglobin A1c levels. An oral glucose tolerance test was performed. Transabdominal or transvaginal ultrasound of the ovaries was done, depending on patients' marital status. All investigations were conducted in the follicular phase of the menstrual cycle. OV >10 mL and/or FNPO ≥12 indicated PCO. A homeostasis model assessment of insulin resistance (IR) value ≥2.6 indicated IR, and metabolic syndrome (MS) was defined according to the international harmonization criteria. Results: Seventy-six participants fulfilled the OV criterion, 70 fulfilled the FNPO criterion, and 89 overall had PCO. Both maximum OV and mean OV had a significant correlation with TT levels (r=0.239, p=0.017 and r=0.280, p=0.005, respectively) and the LH/FSH ratio (r=0.212, p=0.034 and r=0.200, p=0.047, respectively). Mean OV also had a significant correlation with fasting insulin levels (r=0.210, p=0.036). Multivariate binary logistic regression analysis showed that IR (odds ratio [OR], 9.429; 95% confidence interval [CI], 1.701 to 52.271; p=0.010) and MS (OR, 7.952; 95% CI, 1.821 to 34.731; p=0.006) had significant predictive associations with OV alone, even after adjustment for age and body mass index. Conclusion: OV may be more closely related to the androgenic and metabolic characteristics of PCOS than FNPO.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.1
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• pp.31-35
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• 2007

The main purpose of this study was to improve the efficiency and quality of in vitro embryo production in Korean Native Cows (KNC). We examined the effects of ovarian morphologies (Experiment 1) and the culture vessel (Experiment 2) on in vitro maturation (IVM). We measured the subsequent development rates and cell numbers of blastocysts. In Experiment 1, the ovaries of KNC were divided into six groups, based on follicle and corpus luteum (CL) morphology. The development rates to the 2- and 8-cell stages were similar among the six groups. The development rates to blastocyst stages were significantly higher in the group without a CL or follicle (WOCL/F) than in the groups with follicular cysts (FCs), regressive CLs (RCLs) or cystic CLs (CCLs) (p<0.05). The cell number of the inner cell mass (ICM) of blastocysts in the FCs and RCLs groups, and the number of cells in the trophectoderm (TE) in the WOCL/F group, FCs, growing CLs (GCLs) and RCLs were significantly higher than in other groups (p<0.05). The total cell number (TCN) in the WOCL/F, FC and RCL groups was also significantly higher than in other groups (p<0.05). The ICM cell number/TCN ratio was significantly higher in the FC and RCL groups than in the GCL and DF groups (p<0.05). In Experiment 2, oocyte IVM was carried out in culture dishes, in 0.25- or 0.5-ml straws used for freezing sperm. The development rate to the 2-cell stage was significantly higher in the 0.5-ml straw group than in the 0.25-ml straw group. The development rates to the blastocyst stage were similar in the dish and the two straw groups. There were no differences in the cell numbers of ICM, TE or TCN or ICM cell number/TCN ratios between groups.