Current status of assisted reproductive technology in Korea, 2009

Great advances have been made in the field of assisted reproductive technology (ART) since the first in vitro fertilization (IVF) baby was born in Korea in the year of 1985. However, it deserve to say that the invaluable data from fertility centers may serve as a useful source to find out which factors affect successful IVF outcome and to offer applicable information to infertile patients and fertility clinics. This article intended to report the status of ART in 2009 Korean Society of Obstetrics and Gynecology surveyed. The current survey was performed to assess the status and success rate of ART performed in Korea, between January 1 and December 31, 2009. Reporting forms had been sent out to IVF centers via e-mail, and collected by e-mail as well in 2012. With International Committee Monitoring Assisted Reproductive Technologies recommendation, intracytoplasmic sperm injection (ICSI) and non-ICSI cases have been categorized and also IVF-ET cases involving frozen embryo replacement have been surveyed separately. Seventy-four centers have reported the treatment cycles initiated in the year of 2009, and had performed a total of 27,947 cycles of ART treatments. Among a total of 27,947 treatment cycles, IVF and ICSI cases added up to 22,049 (78.9%), with 45.3% IVF without ICSI and 54.7% IVF with ICSI, respectively. Among the IVF and ICSI patients, patients confirmed to have achieved clinical pregnancy was 28.8% per cycle with oocyte retrieval, and 30.9% per cycle with embryo transfer. The most common number of embryos transferred in 2009 is three embryos (40.4%), followed by 2 embryos (28.4%) and a single embryo transferred (13.6%). Among IVF and ICSI cycles that resulted in multiple live births, twin pregnancy rate was 45.3% and triple pregnancy rate was 1.1%. A total of 191 cases of oocyte donation had been performed to result in 25.0% of live birth rate. Meanwhile, a total of 5,619 cases of frozen embryo replacement had been performed with 33.7% of clinical pregnancy rate per cycle with embryo transfer. When comparing with international registry data, clinical pregnancy rate per transfer from fresh IVF cycles including ICSI (34.1%,) was comparable to clinical pregnancy rate per transfer in European Society for Human Reproduction and Embryology report was 32.5% though lower than 45.0% for USA data. There was no remarkable difference in status of assisted reproductive technology in Korea between the current report and the data reported in 2008. The age of women trying to get pregnant was reconfirmed to be the most important factor that may have impact on success of ART treatment.

Integrity of human sperm DNA assessed by the neutral comet assay and its relationship to semen parameters and clinical outcomes for the IVF-ET program / 대한생식의학회지

OBJECTIVE: To explore potential relationships between sperm DNA integrity and both semen parameters and clinical outcomes. METHODS: Semen analysis of 498 samples was performed according to the 2010 criteria of the World Health Organization. The sperm DNA fragmentation Index (DFI) of the semen samples was assessed using a neutral comet assay. RESULTS: Sperm DFI showed a significant correlation with semen parameters, including the patient's age, sperm viability, motility, morphology, and number of leukocytes (p or =14% group (44 cycles, 55.3%), but there was no difference in the other clinical outcomes between the two groups. In the > or =14% group, the pregnancy rates of the ICSI cycles (40.0%) and half-ICSI (44.0%) were higher than conventional IVF cycles (30.7%), but the difference was not statistically significant. CONCLUSION: Along with the conventional semen analysis, the sperm DFI assessed using the comet assay was shown to improve the quality of the semen evaluation. To evaluate the precise effect of ICSI on pregnancy rates in the patients who demonstrate high sperm DFI values, further study is necessary.

Effect of Fragment Removal on Development of Human Fragmented Embryos in IVF-ET Program / 대한불임학회지

OBJECTIVE: To investigate the beneficial effect of fragment removal on the subsequent cell division and clinical outcome of the fragmented human embryos. METHODS: A prospective study was performed in Hanna Women's Clinic and Mizmedi Hospital. Sixty couples undergoing In vitro fertilization-embryo transfer (IVF-ET) program were participated in the present study. The microsurgical fragment removal was performed in 106 fragmented embryos of 29 patients before the transfer. As a control group, 122 fragmented embryos of 31 patients were transferred without the fragment removal. Effects of fragment removal on morphological changes and clinical outcomes of fragmented embryos were investigated. RESULTS: Mean morphological grade (G2.79) of fragmented embryos was significantly improved after the fragment removal (G1.63, p<0.001). Most of the fragmented embryos did not show a regeneration of fragments after the fragment removal during the subsequent development, and a beneficial effect of fragment removal on the development of the fragment removed embryos was observed. Implantation and pregnancy rates of fragment removed embryos were 12.3% and 31.3%, whereas the rates of control group embryos were 6.6% and 22.5%, respectively. There was no statistical significance in the rates between the two groups because of the low number of trials. CONCLUSION: Microsurgical fragment removal improved the subsequent development as well as the morphological grade of fragmented embryos. The fragment removal may be beneficial for neighboring blastomeres by repairing the intercellular communication and removing the secretion of the potential toxic materials by fragments.

Efficient culture system for human embryonic stem cells using autologous human embryonic stem cell-derived feeder cells

Human embryonic stem cells (hESCs) need feeder cells for their maintenance in an undifferentiated state. In conventional culture systems, mouse embryonic fibroblasts (MEFs) serve as feeder cells to maintain hESCs. However, the use of MEFs elevates the risk of transmitting mouse pathogens and thus limits the potential of hESCs in cell replacement therapy. Consequently, the use of human feeder cells would be an important step forward in this in vitro technology. To address this issue, we used fibroblast-like cells differentiated from the Miz-hES6 hESC line (Diff (Miz-hES6)) as feeder cells to support the in vitro growth of three hESC lines. Immunofluorescence microscopy and reverse transcription-PCR assessing the expression of undifferentiated hESC markers revealed all three hESC lines were maintained in an undifferentiated state. In vitro proliferation proceeded as efficiently as when the hESCs were cultured on MEFS. Moreover, karyotype analysis revealed the chromosomal normality of the hESC lines and the Diff (Miz-hES6) feeders themselves after even 50 passages. Furthermore, the hESC lines maintained their pluripotency since they remained capable of forming embryoid bodies (EBs) in vitro. Thus, hESC-derived fibroblast-like cells successfully support in vitro hESC propagation.

Recent Advances in Human Embryonic Stem Cell Research

The 21st century is considered as the era of Biotechnology (BT). Recently, the regenerative medicine using stem cells has been recognized as the future medicine, especially for the devastating diseases such as neurodegenerative diseases, heart disease, diabetes, infertility and liver diseases. Human embryonic stem cells (hESCs) are at the center of the stem cell research due to its ability to proliferate unlimitedly without differentiation (self-renewal) and to differentiate into the derivatives of all three germ layers including germ cells with appropriate treatments (pluripotency). A total of 173 hESC lines have been derived since the first derivation by Thomson et al. in 1998, and 70 hESC lines are currently available for distribution including hESC line (Miz-hES1) established at the MizMedi Hospital. The major goal of hESC research is to provide basic and clinical clues for cell replacement therapy, whose targets are aforementioned incurable diseases. One of the landmarks in hESC research is the derivation of a hESC line from a cloned human blastocyst, which has recently been done by Korean scientists. This made it possible to overcome the issue of immune-mediated rejection following cell replacement therapy using hESCs. Guided differentiation of hESCs into specific cell types by treating growth factors and drugs or by genetic manipulation by using overexpression or an RNAi knockdown system is one of the most active research areas. Combined efforts towards the guided differentiation of hESC into specific cell types and the cloning of hESC from a cloned human blastocyst will overcome a list of diseases hitherto considered to be incurable.

The effect of LH Receptor in the Pregnancy of Poor Responders / 대한불임학회지

OBJECTIVES: To investigate the effect of LH receptor in folliculogenesis, we confirm the expression level of LH receptor (LH-R) mRNA in human granulosa cells (GCs) and its expression levels were analyzed by comparison to embryo developmental rate and pregnancy rate. MATERIALS AND METHODS: GCs were obtained at the time of oocyte retrieval from the patients undergoing IVF-ET program. The patients were divided into two groups: Group I (n=20) is poor responder (retrieved oocyte(s)3ea). After the extraction of total RNA, semiquantitative RT-PCR was performed and the expression level of LH-R mRNA was normalized by beta-actin. Statistical analysis was performed by using Chi(2) test, Student's t-test and Pearson correlation. RESULTS: In Group II, the relative values of LH-R mRNA (0.680 vs. 0.463, p<0.005) and pregnancy rate (54.7% vs. 23.1%, p<0.05) were significantly higher than in Group I. Number of retrieved oocyte(s) was gradually increased when the expression of LH-R mRNA was increased (p<0.05). But the quality of retrieved oocyte and transferred embryo were not related with the expression of LH-R mRNA. When the pregnancy rate was compared with FSH only group and FSH combined with hMG group in the ovarian stimulation protocol, FSH combined with hMG group was significantly higher than FSH only group in Group I (37.5% vs. 0%), and the expression of LH-R mRNA was significantly higher in hMG combined group than FSH only group (p<0.05). CONCLUSION: Expression level of LH-R mRNA has important role in ovarian function related with the response to gonadotrophin in human folliculogenesis. Furthermore these data might provide the evidence that additional use of hMG is helpful to poor responders.

The short-term effects of low-dose estrogen and micronized progesterone on bone turnover markers and serum lipid profiles in postmenopausal women / 대한내과학회지

BACKGROUND: Hormone replacement therapy in postmenopausal women is widely used for the relief of menopausal symptoms and the prevention of bone loss. But, it has been reported that many women discontinue hormone replacement therapy within early period, because the women suffer from breast pain, bleeding and weight gain. Also, further adverse influence of hormone replacement therapy on cardiovascular risk and breast cancer has been suggested. There are many controversies due to conflicting data of that. Recent studies suggest that low-dose estrogen provide bone benefits and micronized progesterone have favorable effects on lipid metabolism and breast density. This clinical trial evaluated the short-term effects of low-dose estrogen and micronized progesterone on bone turnover markers and serum lipid profiles in postmenopausal women. METHODS: This was a 12-week study in which 90 postmenopausal women received hormone replacement therapy. Participants were assigned in equal numbers to the following groups: (1) daily treatment with 0.625 mg conjugated equine estrogens (CEE) with medroxyprogesterone acetate MPA 2.5 mg to 5 mg, daily or cyclically; (2) daily treatment with 0.625 mg CEE with micronized progesterone (MP) 100 mg to 200 mg, daily or cyclically; (3) daily treatment with 0.3 mg CEE with MP 100 mg to 200 mg, daily or cyclically. Changes in bone turnover markers and serum lipid profiles were assessed. RESULTS: At 12-week, all treatment groups significantly improved bone turnover markers and serum lipid profiles, specifically serum alkaline phosphatase, serum osteocalcin, urine deoxypyridinoline and serum high density lipoprotein cholesterol level. CEE 0.625/MPA and CEE 0.625/MP significantly decreased serum low density lipoprotein cholesterol level. CEE 0.625/MPA significantly increased serum triglyceride level. CEE 0.625/MPA produced greater decreases in serum alkaline phosphatase level than CEE 0.625/MP and CEE 0.3/MP. CEE 0.625/MP produced greater increases in serum high density lipoprotein cholesterol level than CEE 0.625/MPA. CEE 0.625/MPA produced greater increase in serum triglyceride level than CEE 0.3/MP. CONCLUSION: Low-dose estrogen and MP generally improved bone turnover markers and serum lipid profiles. But, MP produced lesser favorable effects on a part of bone turnover markers. MP produced significantly greater increases in serum high density lipoprotein cholesterol than that of MPA. And Low-dose estrogen produced significantly lesser increases in triglyceride than that of conventional dose.

The short-term effects of combined hormone replacement therapy and alendronate on bone mineral metabolism in postmenopausal women with osteoporosis / 대한내과학회지

BACKGROUND: The hormone replacement therapy (HRT) and bisphosphonate alendronate are efficacious in the prevention and treatment of postmenopausal osteoporosis. Acting by different mechanisms, these two agents decrease bone resorption and thereby increase or preserve bone mineral density. The combined effects of these medications have not been rigorously studied. This clinical trial examined the combined effects of HRT and oral alendronate on bone mineral density (BMD) and biochemical markers of bone turnover in 50 postmenopausal women with low bone mass. METHODS: This was a 1-year study in which 50 postmenopausal women received daily treatment with 0.625 mg conjugated equine estrogens (with medroxyprogesterone acetate 2.5 mg to 5 mg, daily or cyclically) alone or in combination with 10 mg alendronate. All of the women received a supplement of 600 mg elemental calcium daily. Changes in BMD at the lumbar spine and biochemical markers of bone turnover were assessed. RESULTS: At 1-year, significant increases from baseline in lumbar spine BMD were observed in both treatment groups [combined HRT and alendronate, 10.3%(p < 0.001); HRT alone, 4.5% (p < 0.05)]; Compared with HRT alone, combined HRT and alendronate significantly produced greater increases in BMD of the lumbar spine (p < 0.001). Both treatment groups significantly decreased biochemical markers of bone turnover, especially mean serum osteocalcin level and mean serum alkaline phosphatase level. The combined HRT and alendronate significantly produced greater decreases in mean serum alkaline phosphatase level than HRT alone (p < 0.05). CONCLUSION: Combined use of HRT and alendronate significantly produced greater increases in lumbar spine BMD and greater decreases in biochemical markers of bone turnover than that of HRT alone. Thus, the therapy of combined HRT and alendronate produced favorable effects on bone mineral metabolism in postmenopausal women with osteoporosis.

The short-term effects of combined hormone replacement therapy and alendronate on bone mineral metabolism in postmenopausal women with osteoporosis / 대한내과학회지

BACKGROUND: The hormone replacement therapy (HRT) and bisphosphonate alendronate are efficacious in the prevention and treatment of postmenopausal osteoporosis. Acting by different mechanisms, these two agents decrease bone resorption and thereby increase or preserve bone mineral density. The combined effects of these medications have not been rigorously studied. This clinical trial examined the combined effects of HRT and oral alendronate on bone mineral density (BMD) and biochemical markers of bone turnover in 50 postmenopausal women with low bone mass. METHODS: This was a 1-year study in which 50 postmenopausal women received daily treatment with 0.625 mg conjugated equine estrogens (with medroxyprogesterone acetate 2.5 mg to 5 mg, daily or cyclically) alone or in combination with 10 mg alendronate. All of the women received a supplement of 600 mg elemental calcium daily. Changes in BMD at the lumbar spine and biochemical markers of bone turnover were assessed. RESULTS: At 1-year, significant increases from baseline in lumbar spine BMD were observed in both treatment groups [combined HRT and alendronate, 10.3%(p < 0.001); HRT alone, 4.5% (p < 0.05)]; Compared with HRT alone, combined HRT and alendronate significantly produced greater increases in BMD of the lumbar spine (p < 0.001). Both treatment groups significantly decreased biochemical markers of bone turnover, especially mean serum osteocalcin level and mean serum alkaline phosphatase level. The combined HRT and alendronate significantly produced greater decreases in mean serum alkaline phosphatase level than HRT alone (p < 0.05). CONCLUSION: Combined use of HRT and alendronate significantly produced greater increases in lumbar spine BMD and greater decreases in biochemical markers of bone turnover than that of HRT alone. Thus, the therapy of combined HRT and alendronate produced favorable effects on bone mineral metabolism in postmenopausal women with osteoporosis.

The effect of renal calcium excretion on bone mineral metabolism of postmenopausal women in Korea / 대한내과학회지

BACKGROUND: Although all postmenopausal women are estrogen deficient, women who have postmenopausal osteoporosis may have some defects, in addition to estrogen deficiency, that explain for their higher rates of bone resorption and greater bone loss, relative to those who do not. To test the hypothesis that one of the defects is an impairment in renal calcium conservation, we have investigated relationship between urinary calcium excretion and bone mineral metabolism of postmenopausal women in Korea. METHODS: We have measured 24-hour urinary calcium level; serum osteocalcin level, serum alkaline phosphatase level; urine deoxypyridinoline level; and bone mineral density in 224 early postmenopausal, 158 late postmenopausal and 145 premenopausal women. RESULTS: 33.0 percent (74/224) of early postmenopausal women in Korea had urinary calcium excretion exceeding 4 mg/kg per day. The early postmenopausal women had higher (p < 0.05) values for mean urinary calcium to creatinine ratio of 0.241+/-0.008 mg/mg of creatinine vs. 0.209+/-0.010 mg/mg of creatinine and higher (p < 0.001) mean serum calcium level of 9.15+/-0.02 mg/dL vs. 8.92+/-0.03 mg/dL than the premenopausal women. Hypercalciuric group of early postmenopausal women had higher (p < 0.05) values for mean urine deoxypyridinoline level (8.6+/-0.4 nMol/mMol Vs. 7.7+/-0.2 nMol/mMol), higher (p < 0.05) mean serum alkaline phosphatase level (73.4+/-2.3 U/L Vs. 67.7+/-1.4 U/L) and lower (p < 0.05) mean bone mineral density of femur neck (0.785+/-0.012 g/cm2 Vs. 0.815+/-0.008 g/cm2) than the normocalciuric group. CONCLUSION: The early postmenopausal women had larger numbers of hypercalciuric women and higher values for urinary calcium excretion than the premenopausal women. Hypercalciuric group of early postmenopausal women had higher values for biochemical markers of bone turnover and lower bone mineral density of femur neck than the normocalciuric group. We suggest that hypercalciuria could be accounted for the partial cause of postmenopausal osteoporosis, but further studies are needed to elucidate the direct effect about that.

Effect of Hypoxia on Telomerase Activity in Placental Tissue in Pregnancies with Fetal Growth Restriction / 대한산부인과학회잡지

OBJECTIVE: To observe the difference in telomerase activity (TA) expression in the placenta between fetal growth restriction (FGR) with preeclampsia and those without and to evaluate the effect of oxygen concentration on the TA expression in the trophoblastic cells. METHODS: Telomerase activity was measured in 48 (normal pregnancies, 16; preeclampsia with FGR, 15; normotensive FGR, 17) placentas which were obtained between 32 and 41 weeks' gestations. Trophoblastic cells were extracted from 8 chorionic villi samples obtained from 8-10 weeks' placenta and were cultured in either 2%, 8%, and 20% oxygen atmosphere. Then TA was examined by using telomeric repeat amplification protocol (TRAP) assay. RESULTS: During 3rd trimester of pregnancy, exhibited TA expression in normal pregnancy, FGR complicated by preeclampsia, and normotensive FGR groups were 11 of 16 (68.8%), 4 of 15 (26.7%), and 4 of 17 (23.5%), respectively. Significantly lower level of TA was detected in the FGR group compared to the normal pregnancies (p=0.009), whereas within FGR pregnancies, presence of preeclampsia did not seem to have statistically significant effect on TA expression. TA expression levels were measured by optical density in trophoblasts cultured under various oxygen concentration which revealed that significantly higher TA was exhibited in the cells cultured in 2% oxygen compared to 8% and 20% (p<0.001). However no significant difference was noted in TA between cells cultured in 8% and 20% oxygen. CONCLUSION: Decreased TA in the placenta from pregnancies with FGR was noted regardless of presence of preeclampsia indicating a probable correlation between FGR and placental senescence. Since increased TA was noted in trophoblastic cells that were cultured in hypoxic condition, we could speculate that the intervillous oxygen tension during early-stage placental development plays a certain role in the placental degeneration in pregnancies complicated by FGR and preeclampsia.

The prognostic value of serum CA 125 in endometrial cancer / 대한산부인과학회잡지

OBJECTIVE: The prognostic value of serum CA 125 sampled before the staging operation and the right CA 125 cut-off level in endometrial cancer was evaluated. METHODS: Medical charts of 119 endometrial cancer patients diagnosed and surgically treated at the Department of Obstetrics and Gynecology, Yonsei University College of Medicine from March 1995 to March 2001 were reviewed. The prognostic value of CA 125 was evaluated by comparing it to other established prognostic factors of endometrial cancer such as myometrial invasion, cervix extension, tumor size, and grossly invasive disease. Enzyme immunoassay (Elecsys 2010; Roche Ltd., Basel, Switzerland) was used for the measurement of serum CA 125. RESULTS: The level of presurgically taken serum CA 125 level correlated well with the established prognostic factors of endometrial cancer. CA 125 was one of the information that could be obtained prior to the surgical staging for predicting extensive endometrial cancer, and it could be correlated with the 5 year survival. CA 125 > or = 20 U/ml had a higher sensitivity and lower false negative rate in predicting extensive endometrial cancer compared to the conventionally used level of CA 125> or = 35 U/ml. CONCLUSION: CA 125 has a prognostic value in predicting the extensive endometrial cancer needing lymphadenectomy, therefore it might be included in the baseline study of endometrial cancer for predicting the prognosis and individualizing the treatment modality. CA 125> or = 20 U/ml might be considered as the cut-off level in endometrial cancer for a better sensitivity and false negative rate.

Biochemical Assisted Hatching of Embryos in Human IVF-ET Program / 대한산부인과학회잡지

Human assisted reproductive technology programs have been being developed marvelously during this decade. However, implantation rates following embryo replacement remain low, regardless of increased fertilization rates by ICSI. One proposed possibility limiting the successful implantation is an impaired hatching caused by suboptimal culture conditions. As to improve the hatching potential of blastocysts, assisted hatching by an artificial alteration of zona pellucida(ZP) have been done in many laboratories using the various methods. We tried to investigate whether the supplementation of proteases into culture media has any effect on development, zona structure, and/or hatching of mouse embryos. Supplementation of either pronase E(PE) or proteinase K(PK) in culture media did not affect development up to blastocyst but significantly increased hatching rate. And we observed the alteration of ultrastructure and casein binding properties of ZP in mouse embryos. Also we investigated the effects of protease on development of human embryos and pregnancy rates in human ART program. From July 1994 to December 1996, 792 cycles(for study I) and 1095 cycles(for study II) undergoing the IVF-ET program in MizMedi Hospital were randomly selected for BAH. The concentrations of proteases used in this study were 1microgram/ml PE, 0.1microgram/ml PK and 1microgram/mlPE+0.1microgram/ml PK in HTF with 0.5% human serum albumin(HSA), and in vitro fertilized embryos were cultured for 24 hours. We analyzed the efficiency and stability of biochemical assisted hatching(BAH) according to the clinical profiles of patients and fertilization methods. After cultured in HTF with proteases for 24 hours of human embryos, the thinning in zona pellucida of embryos was observed but its development was not disturbed. Also, clinical pregnancy rates were higher in the PE, PK and PE+PK groups than the control group without proteases(36.0%(32/89), 35.3%(36/102), 35.1%(39/111) vs. 25.5%(125/490), p<0.05). The live birth rate in the PE, PK and PE+PK groups were increased than control, and the abortion rate were not different. They were showed a effect and safety of proteases treatment in human embryos. We selected PE as BAH for study II because of slightly better embryonic morphology and pregnancy rate. In patients over 35 years old, clinical pregnancy rates of the BAH group was higher than that of the control group(31.4%(58/185) vs. 22.2%(51/230); p<0.05). And in the cases with few oocytes retrieval, or less than 3 cycles of IVF-ET, clinical pregnancy rates of the BAH group was significantly higher than that of the control group(36.8%(86/234) vs. 27.2%(93/342), p<0.05; 36.8%(148/402) vs. 29.9%(168/562), p<0.05). In BAH groups, the clinical pregnancy rate was similar between conventional IVF and ICSI group. From above results, it is suggested that improved hatching by protease treatment is due to physiological alteration of ZP structure, giving rise to the similar hatching process to that in vivo. We suggest that BAH using protease is a simple, safe and economic technique compared to the other known assisted hatching techniques in human ART program.

Selective termination in multiple pregnancy guided by transvaginal sonography / 대한산부인과학회잡지

No abstract available.

Transcervical fallopian tube catheterization for proximal tubal obstruction / 대한산부인과학회잡지

No abstract available.

Cytogenetic studies of 384 couples with recurrent abortion / 대한불임학회지

No abstract available.