OBJECTIVE: Evaluation of embryos using early cleavage to 2-cell stage has been proposed, but a critical time-point for selecting embryos is unclear. The aim of the present study is to provide a guideline including critical time-point in the selection of early cleaving embryo for the reduction of multiple pregnancies as well as the increase of pregnancy rate in human IVF. METHODS: This prospective study was performed in 116 cycles from 85 patients who underwent conventional IVF or ICSI at the infertility clinic of Good Moonhwa Hospital from January 2002 to December 2003. Early cleavage (EC) of embryos to 2-cell stage was assessed at 25 h and 27 h postinsemination/microinjection. Embryos that had early cleaved at each time point were designated as EC-1 and EC-2, respectively, while others were designated as non-early cleavage (NEC). RESULTS: At least one early cleavage embryo was observed in 54 (46.6%) for the EC-1 and 84 (72.4%) for the EC-2 of the 116 cycles assessed. Clinical pregnancy rates (PR) were significantly higher in the EC-1 group (66.7%) compared to the EC-2 group (53.6%) or the NEC group (31.2%) (p<0.05). Significant improvement of the pregnancy rate was found when at least two or more embryos were early cleaved at 25 h postinsemination or when the proportion of early cleavage embryo at 25 h postinsemination was higher than 20% (p<0.05). CONCLUSIONS: The critical time-point for the selection of early cleavage embryos with high implantation potential is more effective in 25 h postinsemination/microinjection compared to 27 h. The proportion as well as number of early cleavage embryos is also an important factor for the prediction of pregnancy outcome and the chance of multiple pregnancies. These results demonstrated that the evaluation of early cleavage embryos to 2-cell stage is an easy, simple, and objective method for the selection of good quality embryos suitable for embryo transfer.
Embryo Transfer ; Embryonic Structures* ; Female ; Humans* ; Infertility ; Pregnancy Outcome ; Pregnancy Rate* ; Pregnancy* ; Pregnancy, Multiple ; Prospective Studies ; Sperm Injections, Intracytoplasmic

No abstract available.
Endometrial Neoplasms* ; Female

OBJECTIVE: A number of studies to improve in vitro culture conditions have been tried over past ten years by using co-culture system with helper somatic cells. However, the mechanism of coculture is poorly understood. This study was designed to understand the mechanism for the mode of actual action of co-culture system of ICR strain's 1-cell embryos with human oviduct epithelial cells by examining the effect of conditioned medium and contactless coculture using a cell culture insert on the embryo development and by measuring the level of superoxide anion from conditioned medium after co-culture. METHODS: ICR strain's zygote embryos were cultured in medium alone (control), coculture, conditioned medium, or contactless coculture system for 6 days. Conditioned media (CM) were prepared as following 5 groups. All CM were collected after culturing oviduct cells for 2 days. CM-1 was stored at -20degrees C until use, and CM-2 was prepared just before use as a culture medium. CM-3 was cocultured with embryos and retrieved just before use. CM-4 and CM-5 were derives from the microfilteration of CM-2 and CM-3, respectively, using Microcon-10 (10 kDa molecular weight cut-off). The percentage of the embryos developed to hatched blastocyst stage and the level of superoxide anion in supernatant from medium alone culture (control), coculture, and contactless coculture were measured. RESULTS: The rates of embryo development to the hatched blastocyst stage were significantly higher in coculture (43%) than in control (0%) (p<0.05). The CM-1 group had no embryo development since 2-cell embryonic stage, whereas the CM-2, CM-3, CM-4 and CM-5 groups had the improved development to 4 or 8 cell embryo stage, but the similar rate of development to hatched blastocyst compared to control. The effect of coculture on embryo develpment was disappeared in the contactless coculture group. The level of superoxide anion was significantly reduced in coculture group compared to control. CONCLUSION: It is concluded that the present coculture system overcomes the 2-cell block in vitro and improves the embryo development. This beneficial effect may be due to the direct cell-cell contact between embryo and helper cells or the removal of deleterious components from medium rather than the embryotrophic factors.
Animals ; Blastocyst ; Cell Culture Techniques ; Coculture Techniques* ; Culture Media, Conditioned ; Embryonic Development ; Embryonic Structures ; Epithelial Cells ; Female ; Humans ; Molecular Weight ; Oviducts ; Pregnancy ; Superoxides ; T-Lymphocytes, Helper-Inducer ; Zygote

This study was performed to determine the effects of nitric oxide on human sperm cell function. Semen samples were obtained from normal healthy volunteers. Motile spermatozoas collected by swim-up method were incubated up to 24 hours in Ham's F-10 medium supplemented with a various concentration of sodium nitroprusside (nitric oxide releasing agent). Sperm motility, hyperactivation, acrosome reaction rate, and acrosin activity were determined. The results are as follows; 1. 1mM of SNP resulted in a significant decrease in sperm motility (44.8%+/- 8.9%:78.1%+/-6.3%, and hyperactivation (10.4%+/-6.4%:477%+/-9.5%) after incubation for 3 hours compared with the control group (Ham's F-10 alone), but had no effect on acrosome reaction. 2. At 100muM SNP, sperm motility was reduced after incubation for 6 hours (54.8%+/- 3.2%) compared with that of the control group (82.7% +/- 8.9%), but hyperactivation and acrosome reaction were not affected. 3. However, a lower concentration (less than 101M) of SNP had no effect on sperm motility and hyperactivation for 8 hours of incubation but significantly decreased them when incubation periods were increased up to 24 hours compared with the control group. On the other hand, 1muM and l0muM SNP significantly increased the acrosome reaction rate in both acrosomal status (17.3%+/-5.2%,23.5%+/-4.7%, respectively) and acrosin activity (34.3muIU+/- 10.5muIU, 45.6muIU+/-5.6muIU, respectively) as compared with the control group (7.0%+/-4.0%, 9.5muIU+/-3.4muIU). These results indicate that SNP, NO releasing agent, has a dose-dependent effects on the sperm cell function. Therefore it may positively affect the fertilization by promoting acrosomal reaction at a lower concentration (less than 101M).
Acrosin ; Acrosome Reaction ; Fertilization ; Hand ; Healthy Volunteers ; Humans* ; Nitric Oxide* ; Nitroprusside ; Semen ; Sperm Motility ; Spermatozoa*

Purpose: To evaluate the clinical results of a "Four-in-One" procedure for a habitual dislocation of the patella in children with generalized ligamentous laxity and formation failure of the femoral trochlea. Materials and Methods: Five knees in 4 patients were included in this study. The average age of the patients at the time of surgery was 5.9 years and the subjects were followed up for an average of 41.6 months postoperatively. The clinical results were evaluated using the criteria of the Kujala's scoring system as well as a physical examination and radiological findings. Results: During the follow-up period, there were no recurrent dislocations, knee joint pain, limitations of motion or gait disturbances in any of the cases. The mean Kujala score was 96.8. Two cases had complications related to wound healing but they healed eventually. Conclusion: The "Four-in-One" procedure, which include the lateral retinacular release, medial vector augmentation, semitendinosus tenodesis, and patellar tendon transfer is recommended for a habitual dislocation of the patella in children with generalized ligament laxity and formation failure of the femoral trochlea.
Arthralgia ; Child* ; Dislocations ; Follow-Up Studies ; Gait ; Humans ; Knee ; Knee Dislocation ; Ligaments* ; Patella ; Patellar Dislocation* ; Patellar Ligament ; Physical Examination ; Tenodesis ; Wound Healing

BACKGROUND: Leptin has been found to be involved in the regulation of hematopoiesis processes and angiogenesis. Therefore, we investigated the effect of leptin in the proliferation and angiogenesis of peripheral blood (PB)-derived endothelial progenitor cells (EPCs). METHODS: Mononuclear cells were isolated from PB of healthy male volunteers and were cultured in endothelial cell growth medium-2 (EGM-2). After 6 days of culture, cells were treated with 50 ng/mL vascular endothelial growth factor (VEGF) and/or with various concentrations of leptin (10 ng/mL, 100 ng/mL, 1microgram/mL, and 10 microgram/mL) and were further cultured for one week. Proliferation of EPCs was examined by an assay measuring the uptake of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbo cyanine-labeled acetylated LDL (Dil-ac-LDL) and tubule formation on a matrigel. The control group of cells was not treated with VEGF and/or leptin. RESULTS: The number of Dil-ac-LDL labeled-EPCs, tubule formation on matrigel and the number of cells present along tubules were significantly increased in the leptin-treated groups of cells as compared to the control group or VEGF treated group of cells (P<.05). The effect was synergistically increased in the group of cells co-treated with leptin and VEGF. The number of EPCs was increased in a leptin dose-dependent manner that was maximal at a concentration of 1microgram/mL leptin. CONCLUSION: This study shows that leptin increased in vitro proliferation and angiogenesis of EPCs derived from peripheral blood.
Endothelial Cells ; Hematopoiesis ; Humans ; Leptin* ; Male ; Stem Cells* ; Vascular Endothelial Growth Factor A ; Volunteers

OBJECTIVE: To evaluate the effect of short coasting, by withdrawing both gonadotropins and GnRH agonist (GnRHa), on the prevention in severe ovarian hyperstimulation syndrome (OHSS) without compromising pregnancy outcome. METHOD: Thirty-seven women who had been coasted during COH for IVF were coasted when > or =20 follicles > 15 mm with serum E2 level of 4,000 pg/ml were detected. Coasting was initiated for one or two days depending on the status of follicle on ultrasound and serum E2 level. Both gonadotropin and GnRHa were withheld for coasting. Retrospective study was carried and changes of serum E2 levels, number of oocytes retrieved, fertilization rate, pregnancy rate were compared and analyzed. RESULTS: The mean serum E2 level fell from 6,993 pg/ml on the onset of coasting to 3,396 pg/ml on the day of hCG administration. The mean number of oocytes retrieved and fertilization rate were 15.7 and 70.0%, respectively. Fifteen patients were pregnant (40.6%) and implantation rate was 15.2%. Twenty-six (70.3%) patients were coasted for one day and 11 (29.7%) were coasted for two days. The mean decrease rate of serum E2 level was 43% in one day coasting group and 15% (1st day) and 81% (2nd day) in two day coasting group. The pregnancy outcome was similar between the two groups. After coasting, no severe or moderate OHSS occurred in any patients and mild OHSS occurred in 3 (8.1%) patients. CONCLUSIONS: Coasting for one or two days can be used successfully in the prevention of OHSS without compromising IVF cycle outcome.
Female ; Fertilization ; Gonadotropin-Releasing Hormone* ; Gonadotropins* ; Humans ; Oocytes ; Ovarian Hyperstimulation Syndrome* ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies ; Ultrasonography

OBJECTIVE: To evaluate the effects of serum E2 levels on the day of hCG administration on the pregnancy outcome of IVF-ET after controlled ovarian hyperstimulation (COH). METHODS: Data of 455 cycles of fresh IVF-ET with COH were retrospectively investigated. Serum E2 levels on the day of hCG administration were categorized into 5 groups; A (<1,000 pg/mL), group B (1,000~2,000 pg/mL), group C (2,000~3,000 pg/mL), group D (3,000~4,000 pg/mL), and group E (> or =4,000 pg/mL). RESULTS: Mean E2 levels on the day of hCG administration were 3,745.3 pg/mL and mean number of retrieved oocytes were 10.1. Of 455 cycles, 148 (32.5%) cycles were clinically pregnant. Implantation rate was 12.2% and delivery rate was 18.7%. The number of obtained oocytes increased with increasing levels of serum E2. Pregnancy rate gradually increased as E2 levels increased up to the group D, but began to fall in the group E. In younger women (<38 yrs), the IVF-ET outcomes were similar to those of total patients but in older women (> or =38 yrs), pregnancy rate and delivery rate were significantly higher in the group C than other groups. CONCLUSIONS: This result shows that serum E2 levels have a concentration-dependent effect on the pregnancy outcome and there is an optimal range of E2 levels to achieve for a successful pregnancy. Excessive E2 levels seem more deleterious to the pregnancy outcome in older women aged > or =38 years.
Aged ; Female ; Humans ; Oocytes ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies

OBJECTIVE: Ovarian angiogenesis plays an important role in folliculogenesis. However, little is known about the expression of angiogenic factors during follicular development according to female age. Stromal cell derived factor-1alpha (SDF-1alpha) plays a role in granulosa cell survival and embryo quality as an angiogenic chemokine. Leptin is also involved in folliculogenesis and angiogenesis. This study examined expression of SDF-1alpha and leptin, and their effects on the expression of angiogenic factors in the ovary during follicular development according to female age. METHODS: Ovaries were collected from C57BL mice of two age groups (6-9 weeks and 24-26 weeks) at 6, 12, 24, and 48 hours after 5 IU pregnant mare's serum gonadotropin (PMSG) injection. The expression of ovarian SDF-1alpha and leptin mRNA was evaluated by RT-PCR. In the organ culture experiment, the ovaries were cultured in transwell permeable supports with Waymouth's medium treated with various doses of SDF-1alpha (50-200 ng/mL) or leptin (0.01-1 microg/mL) for 7 days. Then, mRNA expression of vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), and visfatin were examined in the cultured ovaries. RESULTS: Expression of SDF-1alpha and leptin in the ovary was significantly lower in the aged mouse group compared to the young mouse group (p<0.05). Expression of these two factors increased with follicular development after PMSG administration. SDF-1alpha treatment stimulated visfatin expression in a dose-dependent manner, while leptin treatment significantly increased eNOS expression. CONCLUSION: These results suggest that decrease of ovarian SDF-1alpha and leptin expression may be associated with aging-related reduction of ovarian function. SDF-1alpha and leptin may play a role in follicular development by regulating the expression of angiogenic factors in mouse ovaries.
Aged ; Aging ; Angiogenesis Inducing Agents ; Animals ; Chemokine CXCL12 ; Embryonic Structures ; Female ; Gonadotropins ; Granulosa Cells ; Humans ; Leptin ; Mice ; Mice, Inbred C57BL ; Nicotinamide Phosphoribosyltransferase ; Nitric Oxide Synthase Type III ; Organ Culture Techniques ; Ovary ; RNA, Messenger ; Stromal Cells ; Vascular Endothelial Growth Factor A

No abstract available.
Embryonic Development* ; Embryonic Structures* ; Female ; Nitric Oxide* ; Pregnancy