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.

OBJECTIVE: To evaluate the viability and the characteristics of shed endometrial tissues obtained from menstrual fluid during in-vitro culture. METHODS: The menstrual fluids were collected using Wallace catheter from uterine cavity in 10 women with regular menstruation. The menstrual fluids were washed twice, and the pellets, containing blood cells and shed endometrium, were collected and diluted fivefold with Ham's F-10 medium containing 10% fetal bovine serum. The cell suspension was placed on culture dishes, and cultured for 7 days in an incubator. To evaluate the characteristics of the cultured endometrial cells, immunohistochemical (IHC) staining was performed using anti-cytokeratin and anti-vimentin antibody. RESULTS: The mean volume of menstrual fluids and pellets were 0.7ml and 0.3ml, respectively. Only 15% of the shed endometrial tissues were attached and proliferated in culture dishes, which was considered to have viability. Initially, endometrial epithelial cells and fibroblasts were attached and proliferated, and the area of these cells was increased according to prolong the culture time. Stromal cell colonys were located and proliferated on the epithelial cells. IHC staining showed strongly positive for cytokeratin in epithelial cells and for vimentin in stromal cells. In the confocal microscopic observation of 3-dimensional structure of cultured endometrium, cytokeratin-positive cells (epithelial cells) were located in the pheriphery and cytokeratin-negative cells (stromal cells) inside of the structure. CONCLUSION: From our study, shed endometrial tissues in menstrual fluid showed meaningful viability and closed relationship between epithelial cells and stromal cells during in-vitro culture. Thus, we suggest that the in-vitro culture system of shed endometrium is a suitable model for researches of endometriosis.
Blood Cells ; Catheters ; Endometriosis ; Endometrium ; Epithelial Cells ; Female ; Fibroblasts ; Humans ; Incubators ; Keratins ; Menstruation ; Stromal Cells ; Vimentin

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'toysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.
Blastomeres ; Chorionic Villi ; Diagnosis ; DNA ; Dystrophin* ; Embryonic Structures ; Exons ; Genome ; Oocytes ; Polar Bodies ; Spermatozoa

Preimplantation genetic diagnosis (PGD) provides practical option to prevent the termination of pregnancy and miscarriage in couples with high risk of genetic disease or recurrent spontaneous abortion. In balanced chromosomal translocation, PGD can reduce the abortion rate and with PGD for aneuploidy screening, higher implantation rate and lower abortion rate can be obtained in patients with poor reproductive prognosis. Therefore PGD is widely used in ART for improving IVF efficiency. With technical development in single cell, such as FISH, PCR, CGH and microarray, the indications have expanded beyond the monogenic disease and chromosome aberration, as late-onset disease or HLA matching for stem cell donor.
Abortion, Induced ; Abortion, Spontaneous ; Aneuploidy ; Chromosome Aberrations ; Family Characteristics ; Female ; Humans ; Mass Screening ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis* ; Prognosis ; Prostaglandins D ; Stem Cells ; Tissue Donors ; Translocation, Genetic

The aim of this study was to evaluate the efficacy of intravenous immunoglobulin treatment for recurrent spontaneous abortion. Immunologic causes in either alloimmune or autoimmune type have been suggested for more than 80% of these patients. Various immunotherapy including paternal leukocyte transfusion has been used, but there is controversy on its efficacy and side-effects. The proposed immunomodulatory mechanism of intravenous immunoglobulin includes passive transfer of blocking or anti-idiotype antibody, blockade of Fc receptors, enhancement of supressor T-cell function, down regulation of B cell function. In this study, we used intravenous immunoglobulin for the prevention of spontaneous abortion. Five patients with a history of two or more spontaneous abortions were enrolled in this study. Other etiologic factors such as anatomical, chromosomal, hormonal factors were excluded. Three of them were positive for anti-cardiolipin antibody (ACA). When the pregnancy was diagnosed at about five weeks of gestation, 30 g intravenous immunoglobulin was administered and continued up to 28 weeks with three weeks. Ongoing pregnancy beyond 20 weeks was considered successful. Four among five patients (80%) was successful in maintaining pregnancy now ongoing 20, 31, 33, 39 weeks. One patient with ACA positive had abartion due to anembryonic pregnancy. No adverse reaction was observed during the treatment. From these preliminary data, it is suggested that intravenous immunoglobulin treatment may be effective in maintaining pregnancy in patients with unexplained recurrent spontaneous abortion, Further studies are needed to clarify the its immunomodulatory mechanism and establish a more simplified protocol limiting the use at certain critical period of time.
Abortion, Spontaneous* ; Critical Period (Psychology) ; Down-Regulation ; Female ; Humans ; Immunoglobulins* ; Immunotherapy ; Leukocyte Transfusion ; Pregnancy ; Receptors, Fc ; T-Lymphocytes

Preimplantation genetic diagnosis (PGD) is a technique to examine genetic disease or chromosome abnormalities in single cell biopsied from embryos before implantation to uterus. It allows achieving normal pregnancy by transfer of unaffected embryos. The main indications are single gene disorders and recurrent miscarriage related to chromosome aberration and it has advantages to avoid termination of pregnancy or miscarriages in couples with high risk. PGD is also widely applied for aneuploidy screening in assisted reproduction to improve the outcome in infertile patients such as advanced maternal age, although its efficacy still needs to be established. Furthermore, the application of PGD has expanded to other indications, such as late onset-diseases with genetic predisposition and human leukocyte antigen typing for stem cell transplantation. With the advances of molecular diagnostic technologies using single cells, such as fluorescent in situ hybridization, multiplex polymerase chain reaction, fluorescent polymerase chain reaction, linkage analysis, whole genome amplification, array comparative genomic hybridization (array comparative genomic hybridization), and next generation sequencing, PGD can provide more comprehensive and reliable diagnosis.
Abortion, Habitual ; Abortion, Spontaneous ; Aneuploidy ; Chromosome Aberrations ; Comparative Genomic Hybridization ; Diagnosis ; Embryonic Structures ; Family Characteristics ; Female ; Genetic Predisposition to Disease ; Genome ; Humans ; In Situ Hybridization, Fluorescence ; Leukocytes ; Mass Screening ; Maternal Age ; Multiplex Polymerase Chain Reaction ; Pathology, Molecular ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis* ; Prostaglandins D ; Reproduction ; Stem Cell Transplantation ; Uterus

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: To investigate the association between endometriosis and polymorphisms of N-acetyl transferase 2 (NAT2), glutathione S-transferase M1 (GSTM1), and cytochrome P450 (CYP) 1A1 genes in Korean infertile patients. MATERIALS AND METHODS: A total of 303 infertile patients who had undertaken diagnostic laparoscopy during January, 2001 through December, 2003 at Samsung Cheil Hospital enrolled in this study. The patients were grouped according to laparoscopic findings: minimal to mild endometriosis (group I: n=147), moderate to severe endometriosis (group II: n=57), normal pelvic cavity (n=99). Peripheral blood was obtained and genomic DNA was extracted. The genotypes of each genes were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). For NAT2, RFLP was used to detect the wild type (wt) and mutant (mt) alleles, enabling classification into slow (mt/mt) or fast (wt/wt or wt/mt) acetylation genotypes. For GSTM1, PCR was used to distinguish active (+/- or +/+) from null (-/-) genotypes. For CYP1A1, MspI digestion was used to detect the wild type (A1A1), heterozygote (A1A2) or mutant (A2A2) genotypes. RESULTS: The genotype frequencies of NAT2 slow acetylator was 12.8%, 10.9%, 12.8% in group I, group II and control, respectively. The genotype frequencies of GSTM1 null mutation was 55.3%, 41.8%, 53.2% in group I, group II and control, respectively. The genotype frequencies of CYP1A1 MspI polymorphism was 16.3%, 9.1%, 18.1% in group I, group II and control, respectively. No significant difference was observed between endometriosis and normal controls in the genotype frequencies of the NAT2, GSTM1, CYP1A1 MspI polymorphism. CONCLUSION: The NAT2, GSTM1, CYP1A1 gene polymorphism may not be associated with the susceptibility of endometriosis in Korean women.
Acetylation ; Alleles ; Classification ; Cytochrome P-450 CYP1A1 ; Cytochrome P-450 Enzyme System* ; Cytochromes* ; Digestion ; DNA ; Endometriosis* ; Female ; Genotype ; Glutathione Transferase* ; Glutathione* ; Heterozygote ; Humans ; Laparoscopy ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Transferases*

SUMMARY: The present study was carried out to evaluate whether the coculture system of human embryos with Vero cells can improve the quality of embryo or overcome the repetitive implantation failures in order to obtain pregnancy. From January to December 1996, a total 202 cases which patients with the problems of repetitive implantation failures (group I) or those with the poor embryonic quality in their previous cycles (group II) was analysed. The quality of cocultured embryo, pregnancy, on-going and implantation rates between coculture and control groups were compared. Of 93 cases in group I, coculture was performed in 34 cases and conventional IVF for the rest. Of 109 cases in group II, 36 for coculture and 73 for conventional IVF. In group I, pregnancy, on-going and implantation rates in coculture group (14/34 (41.2%), 9/34 (26.5%), 16/81 (19.8%), respectively) were higher than those of control (11/59 (18.6%), 8/59 (13.6%), 12/152 (7.9%), respectively). There is significance in the pregnancy and implantation rates (p=0.028 and p=0.015). In group II, pregnancy, on-going and implantation rates in coculture group (8/36 (22.2%), 5/36 (13.9%), 8/87 (9.2%), respectively) were higher than those of control (5/73 (6.8%), 3/73 (4.1%), 3/158 (1.9%), respectively). Like the result of group 1, there is significance in the pregnancy and implantation rates (p=0.028 and p=0.022). Coculture system with Vero cells works well in the groups of the two indications. Although the case of 3 day-coculture was small as 15 cases in group II, 3 day-coculture improved pregnancy rate (4/15 (26.7%)). Therefore, 3 day-coculture with assisted hatching is recommended to the patients with poor embryonic quality. In conclusion, coculture system with Vero cells can be suggested as an effective method which improves pregnancy rate in those who have repetitive implantation failures or whose embryonic quality was poor in their previous cycles.
Coculture Techniques* ; Embryonic Structures ; Humans ; Humans* ; Pregnancy ; Pregnancy Rate ; Prognosis* ; Vero Cells