The purposes of this study were to evaluate the effects of insulin-like growth factor (IGF)s in peritoneal fluid (PF) from patients with and without endometriosis on the proliferation of endometrial stromal cells and to investigate the effects of type I IGF receptor antibody on the response of endometrial stromal cells to PF from patients with endometriosis. IGFs in PF from patients with endometriosis (n=14) and without endometriosis (n=10) were measured by immunoradiometric assay and PF samples were divided into low IGF-I PF group (less than 85 ng/ml) and high IGF-I PF group (more than 85 ng/ml). Endometrial stromal cells from patients without endometriosis were cultured in serum free media in the presence or absence of 1% PF and thymidine incorporation test were used to evaluate the proliferation of endometrial stromal cells. Also cultures were incubated with type I IGF receptor monoclonal antibody (alpha IR3) before adding PF. PF from patients with endometriosis and without endometriosis increased thymidine incorporation in endometrial stromal cells. In patients with endometriosis, high IGF-I PF group had high IGF-II levels and resulted in higher thymidine incorporation than low IGF-I PE and low IGF-I PF group was noted in patients without endometriosis. There was not a significant correlation between increase in thymidine incorporation and IGF-I levels in PF from patients without endometriosis but in PF from patients with endometriosis. Preincubation with alphaIR3 significantly inhibited the mitogenic response of endometrial stromal cells to PF. Our data indicate that IGF-I in PF may be involved in the growth of ectopic endometrium in patients with endometriosis.
Ascitic Fluid* ; Culture Media, Serum-Free ; Endometriosis* ; Endometrium ; Female ; Humans ; Immunoradiometric Assay ; Insulin-Like Growth Factor I ; Insulin-Like Growth Factor II ; Stromal Cells* ; Thymidine

OBJECTIVE: This study was carried out to evaluate the effect of the isolation methods of inner cell mass from mouse blastocyst, types of feeder cells and treatment time of mitomycin C on the formation rate of ICM colony. METHODS: The inner cells were isolated by conventional immunosurgery, partial trophoblast dissection with syringe needles and whole blastocyst co-culture method. Commercially available STO and primary cultured mouse embryonic fibroblast (pMEF) feeder cells were used, and mitomycin C was treated for 1, 2 or 3 hours, respectively. The formation rate of ICM colony was observed after isolation of ICM and culture of ICM on the feeder cells for 7 days. RESULT: The ICM colony formation rate on STO were significantly higher in partial trophoblast dissection group (58%) than that in immunosurgery (12%) or whole blastocyst culture (16%) group (p<0.05). The formation rate on pMEF feeder layer was higher in partial trophoblast dissection (88%) and whole blastocyst culture (82%) group than that in immunosurgery (16%) group (p<0.05). When mitomycin C treated to pMEF for 2 hours, the formation rate of 88% was significantly higher than those of other conditions. CONCLUSIONS: Above results showed that the efficient isolation method of ICM from blastocyst was the partial trophoblast dissection and the appropriate treatment time of mitomycin C was 2 hours. However, the subculture of ICM colony and characterization of stem cells should be carried out to confirm the efficacy of the partial trophoblast dissection method.
Animals ; Blastocyst ; Coculture Techniques ; Feeder Cells* ; Fibroblasts ; Mice* ; Mitomycin* ; Needles ; Stem Cells ; Syringes ; Trophoblasts

OBJECTIVE: Identification of the regulatory mechanism for arrest and initiation of primordial follicular growth is crucial for female fertility. Previously, we found 15 expressed sequence tags (ESTs) that were specifically abundant in the day-5-subtracted cDNA library and that the B357 clone was novel. The present study was conducted to obtain the whole sequence of the novel gene including B357 and to characterize its mRNA and protein expression in mouse ovary and testis. METHODS: The extended sequence of the 2,965-bp cDNA fragment for the clone B357 was named 5-day-ovary-specific gene-1 (5DOS1) and submitted to GenBank (accession number AY751521). Expression of 5DOS1 was characterized in both female and male gonads at various developmental stages by Northern blotting, real-time RT-PCR, in situ hybridization, Western blotting, and immunohistochemistry. RESULTS: The 5DOS1 transcript was highly expressed in the adult testis, brain, and muscle as compared to the other tissues. In the ovary, the 5DOS1 transcript was detected in all oocytes from primordial to antral follicles, and highly expressed at day 5 after birth and decreased thereafter. In contrast, expression of 5DOS1 showed a gradual increase during testicular development and its expression was limited to various stages of male germ cells except spermatogonia. CONCLUSIONS: This is the first report on the expression and characterization of the 5DOS1 gene in the mouse gonads. Further functional analysis of the 5DOS1 protein will be required to predict its role in gametogenesis.
Adult ; Animals ; Blotting, Northern ; Blotting, Western ; Brain ; Clone Cells ; Cluster Analysis* ; Databases, Nucleic Acid ; DNA, Complementary ; Expressed Sequence Tags ; Female ; Fertility ; Gametogenesis ; Gene Library ; Germ Cells ; Gonads ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Male ; Mice* ; Oocytes ; Ovary* ; Parturition ; RNA, Messenger ; Spermatogonia ; Testis*

OBJECTIVE: We analyzed quantification of mitochondria DNA (mtDNA) to investigate the relationship of mitochondria and pathogenesis of PCOS. MATERIALS AND METHODS: Peripheral blood samples were collected from 28 patients with PCOS who were under the inclusion criteria for PCOS and from 28 healthy controls. Genomic DNA was used to analyze real-time PCR for mtDNA copy number quantification. The mtDNA copy number was compared between the control and PCOS groups. All data was expressed as mean +/- SD. Statistical analysis was assessed by t-test. RESULTS: In this study, the mtDNA CT was 11.67+/-0.422 in PCOS patients and 11.51+/-0.722 in control group, respectively. The mtDNA copy number was 1726410.71+/-407858.591 the patients of in PCOS and 2167887.51+/-252459.28 in control group (p=0.08), respectively. CONCLUSION: In our study, using real-time PCR, there was a tendency of lower mtDNA copy number in the patients of PCOS when comparing to the control group even though statistical difference was not significant. However, more extensive analysis is required to clarity relationship between mtDNA copy number and pathogenesis of PCOS.
DNA ; DNA, Mitochondrial* ; Humans ; Mitochondria ; Polycystic Ovary Syndrome* ; Real-Time Polymerase Chain Reaction

OBJECTIVE: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). METHODS: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test. RESULTS: Mean ages were similar between fresh ET (40.0+/-1.8 years, n=206) and frozen-thawed ET (39.9+/-1.9 years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET (31.2+/-2.3 years, n=40) and frozen-thawed ET (31.9+/-3.1 years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). CONCLUSION: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.
Embryonic Development ; Embryonic Structures ; Female ; Freezing ; Humans ; Insemination ; Pregnancy ; Pregnancy Rate ; Prognosis ; Sperm Injections, Intracytoplasmic ; Uterus

OBJECTIVE: We investigated the expression of uPA and uPAR in eutopic endometrium of advanced stage endometriosis and control patients. METHODS: The 33 endometriosis patients and 32 controls were enrolled. Endometrial samples were obtained from 65 premenopausal women aged 29~44 years, undergoing laparoscopic surgery or hysterectomy for non-malignant lesions. Sufficient samples were collected from 33 patients with endometriosis stage III and IV and 32 controls without endometriosis confirmed by laparoscopic surgery. The mRNA expression of uPA and uPAR from eutopic endometrium were analyzed by RT-QC PCR. RESULTS: The mRNAs of uPA and uPAR were expressed in eutopic endometrium from endometriosis and normal controls throughout the menstrual cycle. Uterine endometrium from women with endometriosis expresses significantly (p<0.05) higher levels of u-PA mRNA than endometrium from normal women without endometriosis in the proliferative phase. There were no significant differences in expression of uPAR in eutopic endometrium between controls and endometriosis patients. CONCLUSION: These results suggest that eutopic endometrium from endometriosis patients may be more invasive and prone to peritoneal implantation because of greater u-PA mRNA expression than endometrium from women without endometriosis. Thus, increased proteolytic activity may be one etiology for the invasive properties of the endometrium resulting in the development of endometriosis.
Endometriosis* ; Endometrium* ; Female ; Humans ; Hysterectomy ; Laparoscopy ; Menstrual Cycle ; Polymerase Chain Reaction ; Proteolysis ; RNA, Messenger* ; Urokinase-Type Plasminogen Activator

OBJECTIVE: To evaluate the impact of endometriosis on IVF-ET cycles and to compare IVF outcomes between stage I/II and stage III/IV endometriosis. METHODS: We analyzed 697 patients (1,199 cycles) with endometriosis (stage I-II: 638 cycles, stage III-IV: 561 cycles) and 325 pts (459 cycles) with tubal factor as controls between January 1994 and April 2004. Pts with endometriosis were diagnosed by laparoscopy and medical and surgical treatment were done in 353 cycles (55.3%) and 466 cycles (83.1%) of stage I-II/stage III-IV endometriosis. Cycles with age>35 years or FSH>20 mIU/mL or severe male factor infertility were excluded. RESULTS: The number of retrieved oocytes (9.97+/-7.2 vs. 13.4+/-7.9 (p<0.0001)), total number of embryos (6.5+/-4.8 vs. 9.1+/-5.6 (p<0.0001)), and good quality embryos (2.43+/-1.6 vs. 2.74+/-1.7 (p=0.013)) significantly decreased in stage III-IV endometriosis than in control. But pregnancy rate of stage III-IV endometriosis was comparable with control (35.7% vs. 36.8%). Fertilization rate and number of total embryos were lower in stage I-II endometriosis than in control (64.8+/-22.9 vs. 70.8+/-20.8 (p<0.0001), 7.6+/-5.0 vs. 9.1+/-5.6 (p<0.0001)). In patients with medical and surgical treatment of endometriosis, pregnancy rate and live birth rate was significantly lower in stage I-II than in stage III-IV endometriosis (29.2 vs. 36.2 (%), p=0.045, 23.9 vs. 31.5 (%), p=0.043). There was no difference in the mean age, but the duration of infertility was significantly longer (56.5+/-26.3 vs. 46.9+/-25.8 (mon), p<0.0001) and fertilization rate was lower (64.7+/-23.3 vs. 70.5+/-22.7 (%), p=0.001) in stage I-II than stage III-IV endometriosis. CONCLUSION: We suggest that IVF should be considered earlier in patients with minimal to mild endometriosis because of significantly decreased fertilization rates.
Embryonic Structures ; Endometriosis* ; Female ; Fertilization ; Humans ; Infertility ; Laparoscopy ; Live Birth ; Male ; Oocytes ; Pregnancy Rate

No abstract available.
Counseling* ; Infertility*

No abstract available.
Abortion, Spontaneous* ; Classification* ; Female ; Humans ; Pregnancy

No abstract available.
Humans ; Male*