Efficacy of corifollitropin alfa followed by recombinant follicle-stimulating hormone in a gonadotropin-releasing hormone antagonist protocol for Korean women undergoing assisted reproduction / 대한생식의학회지

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.

Comparison of Clinical Efficacy between a Single Administration of Long-Acting Gonadotrophin-Releasing Hormone Agonist (GnRHa) and Daily Administrations of Short-Acting GnRHa in In Vitro Fertilization-Embryo Transfer Cycles

This study was aimed to evaluate the efficacy of a single administration of long-acting gonadotrophin-releasing hormone agonist (GnRHa) as compared with daily administrations of short-acting GnRHa in controlled ovarian hyperstimulation (COH) for in vitro fertilization and embryo transfer (IVF-ET) cycles. The mean dosage of recombinant follicle-stimulating hormone (rFSH) required for COH (2,354.5+/-244.2 vs. 2,012.5+/-626.1 IU) and the rFSH dosage per retrieved oocyte (336.7+/-230.4 vs. 292.1+/-540.4 IU) were significantly higher in the long-acting GnRHa group (N= 22) than those in the short-acting GnRHa group (N=28) (p<0.05). However, the mean number of visit to the hospital that was required before ovum pick-up (3.3+/-0.5 vs. 22.2+/-2.0) and the frequency of injecting GnRHa and rFSH (12.8+/-1.2 vs. 33.5+/- 3.5) were significantly decreased in the long-acting GnRHa group (p<0.0001). The clinical pregnancy rate, implantation rate, and early pregnancy loss rate were not significantly different between the 2 groups. So, we suggest that a single administration of long-acting GnRHa is a useful alternative for improving patient's convenience with clinical outcomes comparable to daily administrations of short-acting GnRHa in COH for IVF-ET cycles.

Efficiency of Percoll Gradient, PureSperm Gradient, and Swim-up Separation Techniques in Normal Semen Samples / 대한남성과학회지

PURPOSE: The aim of this study was to compare sperm concentration, motility, viability, and morphology using Percoll gradient, PureSperm gradient, and the swim-up method in normal semen samples. MATERIALS AND METHODS: Ten normal semen samples were divided into three fractions. Motile sperm were isolated with Percoll gradient, PureSperm gradient, and swim-up method. Sperm concentration, motility, viability, and morphology were determined before and after separation. RESULTS: The sperm concentrations were not significantly different among the three METHODS: Percoll gradient(34.3+/-9.4x10(6)/ml), PureSperm gradient(37.6+/-16.6x10(6)/ml), and swim-up(27.3+/-6.4x106/ml). There was no significant difference in sperm motility among the three METHODS: Percoll gradient(93.5+/-1.6%), PureSperm gradient(92.7+/-4.4%), and swim-up(95.7+/-2.7%). When sperm motility was measured 24 hr later, the results were similar among the three METHODS: Percoll gradient(81.7+/-15.5%), PureSperm gradient(84.3+/-12.2%), and swim-up(89.4+/-5.1%). Sperm viability and morphology were slightly higher in swim-up method than the other methods, but the differences were not statistically significant. Sperm viability datas were: Percoll gradient(85.5+/-5.5%), PureSperm gradient(85.6+/-3.7%), and swim-up(88.6+/-6.6%). Morphology datas were: Percoll gradient(82.0+/-10.7%), PureSperm gradient(73.9+/-9.3%), and swim-up(83.7+/-8.5%). CONCLUSIONS: The swim-up method resulted in viability and morphology that were slightly higher than the other methods. However, all methods were useful for sperm preparation in normal semen samples.

Pregnancy Outcomes after Transfer of Frozen-thawed Embryos following ICSI with Ejaculated, Fresh and Frozen-thawed Testicular Sperm / 대한산부인과학회잡지

OBJECTIVES: This study was performed to evaluate the pregnancy rate following the transfers of frozen- thawed embryos which was derived from intracytoplasmic sperm injection (ICSI) using sperm obtained by ejaculated, testicular sperm extraction (TESE), and frozen-thawed testicular sperm extraction (t-TESE). METHODS: Frozen-thawed embryos were successfully transferred to the patients in 664 cycles among 695 cycles from January 1998 to December 2002, where ICSI was done with various origins of sperm. Subjects were divided into three groups according to the origin of sperm; ejaculated sperm group as a control (n=535), TESE group (n=98) and t-TESE group (n=62). After conventional ICSI, the supernumerary PN stage or developing embryos were cryopreserved by slow freezing protocol with 1, 2-propanediol as cryoprotectant. RESULTS: The survival rate of frozen-thawed embryos was 77.7% (2515/3236) in ejaculated sperm group, 76.6% (441/576) in TESE group and 83.9% (292/348) in frozen-thawed TESE group, respectively. The difference of survival rate of between t-TESE group and other two groups was statistically significant (p<0.01). The good embryo formation rate and positive beta-hCG rate was 46.3% (1164/2515), 28.8% (148/513) in ejaculated sperm group, 49.2% (217/441), 36.6% (34/93) in TESE group and 46.2% (135/292), 34.9% (22/63) in frozen-thawed TESE group, respectively. CONCLUSION: This study demonstrates that comparable pregnancy rate and implantation rate could be achieved after the transfer of frozen-thawed embryos following ICSI using various sources of sperm. As there was no statistically significant difference in pregnacy rate between ICSI with fresh testicular sperm and with frozen-thawed testicular sperm, the sequential cryopreservation of supernumerary testicular sperm and embryos may be a useful method for increasing pregnancy outcome in infertile couples with male factor.

Light and Electron Microscopic Observation in the Frozen-thawed Mouse Testicular Tissues / 대한불임학회지

OBJECTIVE: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. METHODS: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. RESULTS: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. CONCLUSIONS: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

Light and Electron Microscopic Observation in the Frozen-thawed Mouse Testicular Tissues / 대한불임학회지

OBJECTIVE: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. METHODS: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. RESULTS: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. CONCLUSIONS: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

Comparative Analysis of Pregnancy Outcomes after In Vitro Fertilization with Intracytoplasmic Sperm Injection (IVF-ICSI) between Obstructive and Non-obstructive Azoospermia / 대한불임학회지

OBJECTIVE: To compare the pregnancy outcomes after in vitro fertilization with intracytoplasmic sperm injection (IVF-ICSI) between obstrucvtive and non-obstrucvtive azoospermia. METHODS: From January 1994 to December 2002, 524 patients with obstructive azoospermia (886 cycles) and 163 patients with non-obstructive azoospermia (277 cycles) were included in this study. Microsurgical epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE) in obstructive azoospermia and TESE in non-obstructive azoospermia were perfomed to retrieve sperm, which was used for ICSI and then fertilized embryos were transferred. The results of ICSI-fertlization rate (FR), clinical pregnancy rate (CPR), clinical abortion rate (CAR) and delivery rate (DR)- were statistically analysed in obstructive versus non-obstructive azoospermia. RESULTS: There were no differences in the number of retrieved oocytes, injected oocytes for ICSI and oocyte maturation rate. FR was significantly higher in obstructive than non-obstructive azoospermia (71.7% vs. 61.1%, p<0.001). There was no difference in CPR per embryo transfer cycle. After pregnancy was established, however, CAR was significantly higher in non-obstructive than obstructive azoospermia (25.6% vs. 12.5%, p=0.004). DR per clinical pregnancy cycle was significantly higher in obstructive than non-obstructive azoospermia (78.0% vs. 64.4%, p=0.012). In the karyotype ananlysis of abortus, abnormal karyotypes were found in 75.0% (6/8) of obstructive and 55.6% (5/9) of non-obstructive azoospermia. CONCLUSION: Our data show significantly higher FR in obstructive than non-obstructive azoospermia. Though there was no differrence in CPR, CAR was significantly higher in non-obstructive than obstructive azoospermia. The abortion may be related to the abnormal karyotype of embryo, but further investigations are necessary to elucidate the cause of clinical abortion in azoospermia.

Vitrification of Mouse Blastocyst Using Cryoloop / 대한불임학회지

OBJECTIVE: The aim of this study is to compare the efficiency of a method for the cryopreservation of mouse blastocyst. METHODS: Mouse embryos were obtained at 2-cell stage and cultured to blastocyst stage in T6 medium supplemented with 10% fetal bovine serum. Morphologically normal blastocysts were collected and randomly divided to one control and four experimental groups. In control group, blastocysts were cultured in vitro continuously for additional two days. In group 2, blastocysts were exposed to vitrification solution (ethylene glycol) only without cryopreservation (exposure only group). In group 3, 4 and 5, blastocysts were cryopreserved by slow-freezing procedure with glycerol (slow-freezing group) or by vitrification procedure using EM grids (EM grids group) and cryoloop (cryoloop group), respectively. Frozen blastocysts were thawed and cultured for additional two days. Twenty four hours after thawing, some blastocysts were fixed and stained with Hoechst 33342 (bisbenzimide) and the number of nuclei in each blastocysts were counted to confirm the survival of blastocysts in experimental groups. RESULTS: Survival rate and hatching rate of the blastocysts in slow-freezing group (24 h: 72.4% and 66.0%, 48 h: 63.2% and 64.6%) and EM grids group (24 h: survival rate 77.3%, 48 h: 70.1% and 71.4%) were significantly lower (X2-test p<0.05) than those of control group (24 h: 93.4% and 86.0%, 48 h: 88.5% and 90.7%). In contrast, the survival rate and hatching rate of the blastocysts in cryoloop group (24 h: 84.1% and 84.1%, 48 h 79.3% and 87.7%) is well compared with those in the control group. The mean (+/-SD) cell number of blastocyst in the exposure only (89.2+/-11.5), EM grids (85.0+/-10.3) and cryoloop (89.0+/-11.0) groups, except slow-freezing group (79.0+/-10.0), were not significantly different from that of control group (93.1+/-13.9) 24 h after thawing (Student's t-test). CONCLUSION: This study demonstrates that higher survival rate of vitrified-thawed mouse blastocyst can be obtained using cryoloop as the embryo container at freezing rather than slow-freezing or vitrification using EM grids. The results of this study suggest that vitrification using cryoloop (with ethylene glycol) may be a preferable procedure for mouse blastocyst cryopreservation and could be applied to the human blastocyst cryopreservation.

Influence on the Pregnancy Rate of Motility of Fresh and Frozen Testicular Spermatozoa in Obstructive Azoospermic Patients / 대한불임학회지

OBJECTIVE: ICSI with testicular sperm could achieve optimal fertilization and pregnancy. This study was performed to observe the influence on fertilization and pregnancy of motility of fresh testicular sperm and sperm extracted from frozen-thawed seminiferous tubules in obstructive azoospermia. MATERIALS ANDMETHODS: We analysed clinical outcome of ICSI using fresh testicular sperm and sperm extracted from thawed seminiferous tubules. The presence of motility were compared to determine the factor for optimal fertilization and pregnancy rates. RESULTS: In 316 cases of TESE-ICSI in obstructive azoospermia, ICSI with fresh testicular sperm (fresh sperm group) were 163 cases and ICSI with sperm testicular sperm extracted from frozen-thawed seminiferous tubule (thawed sperm group) were 153 cases. The fertilization rates were 71.3% and pregnancy rates were 32.5% in fresh sperm group, in thawed sperm group, 65.1% and 33.3% respectively. The fertilization and pregnancy rates of motile and non-motile testicular sperm were 72.9% and 33.6%, 50.0% and 18.2%, respectively (p<0.05). The fertilization and pregnancy rates of motile and non-motile sperm extracted from the thawed seminiferous tubule were 67.8% and 34.7%, 55.1% and 28.1%, respectively (p<0.05). The comparative of the results of ICSI using motile fresh testicular sperm and motile sperm extracted from thawed seminiferous tubule, fertilization and pregnancy rates were not significantly different (72.9% and 33.6%, 67.8% and 34.7%, respectively). CONCLUSION: These results suggest that successful pregnancy in TESE-ICSI treatment is influenced by the motility of fresh testicular sperm and sperm extracted from thawed seminiferous tubule in obstructive azoospermic patients.

Effect of Hypo-osmotic Swelling (HOS) Test on Subsequent Post-thaw Testicular Spermatozoa / 대한불임학회지

OBJECTIVES: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. MATERIALS AND METHODS: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from LN2 and letting them at room temperature for 10 min followed by 37degrees C water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. RESULTS: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospermic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. CONCLUSIONS: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.

Analysis of Influencing Factors on the Successful Pregnancy in Human IVF-ET Program Using Testicular Sperm Extraction (TESE) with Intracytoplasmic Sperm Injection (ICSI) / 대한비뇨기과학회지

No abstract available.

Adverse Effect of Human Hydrosalpingeal Fluid on the Development of Mouse Embryo (II) / 대한불임학회지

In our previous study, we observed that hydrosalpingeal fluid (HSF) adversely effect mouswe embryo development and hatching. The aim of this study was to evaluate the effect of HSF as assessed by the blastocyst development rate (BDR) and by cell counting in vitro HSF was collected from nine patients undergoing salpingoneostomy to correct hydrosalpinx. Two-cell embryos were obtained from superovulated ICR mice. T6 medium and T6+/-0.4% bovine serum albumin were used as control media. T6 medium containing 10% or 50% HSF and 100% HSF from each patient were used as test media. Nine to 15 embryos were cultured in microdrops prepared from each of these media. To assess the total cell number within each blastocyst, the blastocysts were fixed and stained with Hoechst 33342 to facilitate cell counting. The mean BDR in two control media were 88.89% and 85.40%. The mean BDR in media containing 10%, 50%, 100% HSF were 85.87%, 89.58% and 75.57%, respectively (*: p<0.05). The overall mean cell count (+/-SEM) in control media were 87.6+/-9.65 and 90.12+/-11.38. The BDR was affected adversely only by 100% HSF and not in media containing 10% or 50% HSF. Mean cell counts were decreased significantly only in blastocysts cultured 100% HSF (63.8+/-13.66; p<0.01) but not in blastocysts cultured in 10% or 50% HSF (91.3+/-12.44 and 82.9+/-18.27, respectively). Thus, it is concluded that HSF has no embyotoxic effect but has a mildly negatively effect on embryonic growth and development.

Significance of the CT in rectal cancer

No abstract available.

A clinical review of snake bite

No abstract available.