• Reproductive and Developmental Biology
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• 제34권3호
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• pp.169-173
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• 2010

Correlations between cumulus cells and germinal vesicle (GV) chromatin configuration were examined in porcine oocytes. Cumulus-oocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three categories according to cumulus cell morphology. "A" group was compacted COCs with more than three cumulus cell layers. "B" group was COCs with less cumulus cell layers than "A" group. "C" group was COCs with one or less layer of cumulus cells. Cumulus cells were removed 0.1% hyaluronidase, and denuded oocytes were stained with Hoechst 33342. GV chromatin configuration was classified into GV-Con and GV-Dis. GV-Con meant that a nucleus was surrounded by condensed chromatin in a ring. GV-Dis meant that filamentous chromatin clumps were distributed in nucleus. The proportion (80.2%) of GV-Con in "A" group was significantly higher than "B" (62.0%) or "C" (44.9%). The proportion (55.1%) of GV-Dis in "C" group was significantly higher than "A" (19.8%) or "B" (38.0%). The meiotic competence of COCs was examined after 44 h culture. The proportion (90.0%) of oocytes reaching to metaphase II (M-II) in "A" group was significantly higher than "B" (76.5%) or "C" (45.5%). In conclusion, oocytes with good quality cumulus cell layers are synchronized early GV stage, and early GV stage is important for meiotic competence in pigs.

• 대한한방부인과학회지
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• 제23권4호
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• pp.67-79
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• 2010

Purpose: To evaluate the effect of Korean medical treatment as supplementary treatment method in ART. Methods: The research was investigated with Korean, Chinese and western medical clinical journals of ART with herbal medicine, acupuncture, and other oriental medical treatment as supplementary treatment. Results: ART with additional herbal treatment achieved notably better result when ovulation rate, quality of oocyte, pregnant rate were compared. In the indepth review and analysis of five journals of ART with acupuncture intervention, firm effective results were not found, however, acupuncture intervention on patients with embryo transplantation had higher achievement rate in artificial insemination. Conclusions: According to the review of the Korean, Chinese, and western medical journals of the effect of Korean medical treatment as supplementary treatment method in ART, acupuncture, herbal medicine, and other oriental medical therapies in ART contribute to the improvement of the outcome of ART. The limit in the research was limited strong enough well-designed RCT studies as evidence for they were very seldom.

• 한국발생생물학회지:발생과생식
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• 제14권1호
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• pp.27-34
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• 2010

A fertilized oocyte can get the competence for implantation through cleavage and stage-specific gene expression. These are under the control of autonomous and exogenous regulators including physiological culture condition. Endogenous and exogenous growth factors are considered as critical regulators of cleaving embryos during travel the oviduct and uterus. In this study, an effort was made to evaluate comprehensively the quality of embryos for implantation, grown in media enriched with EGF and PAF. The study evaluated developmental rates on given time, blastulation and hatching rates, and adhesion rates. Developmental rates of blastocyst to the hatching stage were significantly high in PAF treated group compared to the control in a dose-dependent manner but not in EGF group. Implantation rates were significantly high both PAF and EGF in a dose-dependent manner. H7, a PKC inhibitor, blocked the process of hatching of the blastocysts but combined treatment of EGF and PAF enhanced the hatching and implantation of blastocsyts. Based on these results it is suggested that EGF and PAF support acquirement of implantation competence at blastocyst stage through a PKC pathway.

• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.52-55
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• 2017

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.407-414
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• 2011

The development of embryos reconstructed by somatic cell nuclear transfer (SCNT) is dependent upon numerous factors. Central to development is the quality and developmental competence of the recipient cytoplast and the type of the donor nucleus. Typically metaphase of the second meiotic division (MII) has become the cytoplast of choice. Production of a cytoplast requires removal of the recipient genetic material, however, it may remove proteins which are essential for development or reduce the levels of cytoplasmic proteins to influence subsequent reprogramming of the donor nucleus. In this study, enucleation at MII did not affect the activities of either MPF or MAPK kinases. Immunocytochemical staining showed that both Cyclin B1 (MPF) and Erk1/2 (MAPK) were associated with the meiotic spindle of AI/TI oocytes with little staining in the cytoplasm, however, at MII association of both proteins with the spindle had reduced and a greater degree of cytoplasmic distribution was observed. The analysis of oocyte proteins removed during enucleation is a difficult approach to the identification of factors which may be depleted in the cytoplast. This is primarily due to the large numbers of aspirated karyoplasts which would be required for the analysis.

• Clinical and Experimental Reproductive Medicine
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• 제45권4호
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• pp.189-194
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• 2018

Objective: The aim of this study was to compare the rate of maturation, fertilization, and embryo development of in vitro-matured human oocytes derived from pregnant and non-pregnant women. Methods: Immature oocytes were obtained by needle aspiration from 49 pregnant women (group 1) who underwent a cesarean section at term and 77 non-pregnant women (group 2) who underwent a gynecological operation during the same period (8 months). Healthy immature oocytes (530 in group 1 and 539 in group 2) were cultured and assessed for maturation 36 hours later. Mature oocytes were inseminated by intracytoplasmic sperm injection and cultured up to 144 hours. Results: The percentage of degenerated oocytes was significantly higher (12.1% vs. 6.3%; p<0.001) in group 1 than in group 2. There was no significant difference in the maturation rate (66.8% vs. 68.1%; p=0.698), fertilization rate (66.7% vs. 67.6%; p=0.857), or the rate of formation of good-quality blastocysts (46.2% vs. 47.2%; p=0.898) in oocytes obtained from pregnant and non-pregnant women. Conclusion: The developmental competence of immature oocytes did not differ between pregnant and non-pregnant women.

• Clinical and Experimental Reproductive Medicine
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• 제28권4호
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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.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.71-77
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• 2012

The objective of this study was to examine the effect of thymidine treatment during $in$ $vitro$ maturation (IVM) of porcine follicular oocytes on blastocyst development. Porcine oocytes were treated with thymidine (10 mM, 20 mM and 30 mM) for 2 or 6 hr in the preiods of IVM I and/or II. The survival rates of the blastocysts in the 6 hr treatment groups of 10 mM and 20 mM during IVM I period were significantly higher than those of control group ($p$<0.05). However, the survival rate of the blastocysts in the 2 hr treatment group of 20 mM during IVM II period was significantly higher than control group ($p$<0.05). Furthermore, the survival rate of the blastocysts in the 6 hr treatment group of 30 mM during IVM II period was significantly lower than control group ($p$<0.05). Consistent with the previous result, blastocyst development of both IVM I and II treatment group was also showed as similar pattern. Total and apoptotic cell numbers of blastocysts derived from thymidine treated porcine oocytes were examined by using Tunel assay. The results showed that there was no significant differences in total cell number of blastocysts between thymidine treated and untreated groups. However, apoptosis-positive cells in the thymidine treated group (6 hr IVM I) were significantly lower than those of other groups ($p$<0.05). Taken together, these results indicate that high quality oocytes were selected by DNA synthesis mechanism according to high concentration thymidine treatment during porcine oocyte maturation. Therefore, we concluded that presumptive selected oocytes by thymidine treatment during maturation periods improved the further embryo development and embryonic quality of IVF embryos by decreasing the incidence of apoptosis in preimplantation porcine embryos.

• 대한한방부인과학회지
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• 제31권4호
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• pp.89-109
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• 2018

Objectives: The purpose of this systematic review is to assess the effects of herbal medicine with in vitro fertilization-embryo transfer (IVF-ET) based on the current evidence. Methods: Eligible randomized controlled trials (RCT) searched from Pubmed which compared a combination of herbal medicine and IVF with IVF alone were included. Results: Sixteen trials, in which 2025 women involved, were included in this review. The review results showed that the effect of traditional herbal medicine can improve the clinical pregnancy rate (herbal medicine intervention: 30.36~63.64%, Control: 9.38~47.5%) and rate of high quality oocytes and embryos. The mechanism may be through regulating follicular fluid to improve microenvironment for oocytes which would lead to a successful embryo implantation. Conclusions: This analysis showed that combination of IVF and traditional herbal medicine used in the included trials improve clinical pregnancy rate and IVF success. During in vitro fertilization, TCM can regulate the microenvironment in the follicular fluid to mature the oocyte, improve the quality of the embryo, and help the development and implantation of the embryo. Further large randomized placebo controlled trials are needed to confirm the effectiveness of traditional herbal medicine with concurrent IVF.

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.75-81
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• 2000

Objective: The objective of this study is to influence of sequential embryo transfers in an invitro fertilization was examined. Method: After in vitro fertilization, a maximum of 6 fertilized oocytes was enrolled in this study. At day 3 after an oocytes retrieval, embryos with good quality were transferred (mean 4.9), remaining embryos (mean 2.0/cycle) were cryopreserved at blastocyst stage (Group 1). At day 5 after oocytes collection, second a embryo transfer (mean 1.2/cycle) was performed, if one of these embryos had reached the blastocyst stage (Group 2) using P1 supplemented with 10 SSS and 30% Follicular fluid. No statistical difference in the pregnancy rate could be seen between the group without a second embryo transfer (n=21; 28.6%) and the group with a second transfer (n=52; 28.8%). Results: The incidence of multiple pregnancy rate per embryo transfer was not statistically different between both group and no high-rank multiple pregnancy (greater than triplete) were observed (0.9%, 15.4%, respectively, p=0.74, ${\chi}^2$). Out of 114 cycles (506 embryos) cultured embryos in group 2, 52 cycles (159 embryos, 29.8%) reached the blastocyst stage. Conclusion: The second transfer did not have a significant effect on the pregnancy rate. The most important factor for the pregnancy seems to be the quality of the embryos transferred on day 3 following oocyte retrieval. We recommend embryo transfer is performed only one, day $2{\sim}3$ or D5.