https://www.jri.ir/ "Journal of Reproduction & Infertility" is owned, published, and copyrighted by ©2009 Avicenna Research Institute. No parts of this journal may be reproduced in any form or by any means unless properly referenced or sent a notification letter through www.jri.ir en Isolation, Culture and Characterization of Human Sertoli Cells by Flow Cytometry: Development of Procedure <p>Background: The sertoli cells in the testis create unique and safe environment to protect seminiferous tubules from auto antigens and invading pathogens. These cells produce the survival factor of the blood-testis barrier and produce special materials such as androgen binding proteins and contribute to the coordinated action of spermatogenesis. Given that the sertoli cells play an essential role in spermatogenesis and the lack of these cells leads to the disruption of spermatogenesis, it is necessary to investigate the behavior and performance of these cells. To achieve this goal, the cells must first be extracted. The aim of this study was to develop a procedure to isolate, culture, and characterize human sertoli cells.<br /> Methods: In order to isolate the sertoli cells of azoospermia patients who underwent (testicular sperm extraction) TESE surgery, washing up and multi_stage enzyme digestion of single cells, culture on petri dishes impregnated with datura stramonium lectin agglutinin (DSA) were done and then the cells were passaged for several times and isolated. For more purification, flow cytometry method with FSH receptor antibody was used. Immunocytochemistry assays and Elisa test for identification of these cells were employed.<br /> Results: The purification method resulted in more than 97% purity. The nature of sertoli cells was confirmed by morphology evaluation, detecting anti-mullerian hormone in sertoli cell culture media and the presence of FSH receptor on sertoli cells.<br /> Conclusion: This study introduced and applied a method to isolate, culture, and purify human sertoli cells with high purity which made possible further researches on these cells.</p> https://www.jri.ir/article/696 The Relationship Between Uterine Artery two-dimensional Color Doppler Measurement and Pregnancy Outcome: A Prospective Observational Study <p>Background: The purpose of the study was to evaluate the role of uterine blood flow parameters measured by uterine artery two-dimensional (2D)-power color doppler (PCD) ultrasound in predicting fertility outcomes in women undergoing IVF-ET cycles.<br /> Methods: In this prospective observational study, a total of 188 infertile women who underwent IVF-ET cycles were investigated. Uterine artery 2D-PD measurements were taken during early follicular phase and on day of trigger. Pulsatility Index (PI), Resistant Index (RI), Peak Systolic Velocity (PSV), and Systolic/Diastolic ratio (S/D) were measured. Statistical correlation was sought between the doppler parameters and fertility outcomes.<br /> Results: The pregnancy rate was 40.43% (76/188). The women who conceived (n=76) (Group A) were found to have mean age of 31.2&plusmn;3.9 years whereas the non-pregnant group of women (n=112) (Group B) had mean age of 31.45&plusmn;4.25 years. The mean PI measurements subsequently during early follicular phase and on the day of hCG trigger between group A and group B were comparable (2.09&plusmn;1.15 versus 1.9&plusmn;0.95; p=0.385 and 1.86&plusmn;1.12 versus 2.03&plusmn;1.0; p=0.192, respectively). No significant changes in the uterine artery PSV values and S/D values and RI were noted during the cycle.<br /> Conclusion: Uterine artery doppler evaluation in women undergoing IVF cycles was not predictive of the pregnancy outcomes.</p> https://www.jri.ir/article/700 Antimicrobial Properties of Amniotic and Chorionic Membranes: A Comparative Study of Two Human Fetal Sacs <p>Background: There is evidence of antibacterial properties of human chorioamniotic layer. However, the distinctive contribution of its individual parts, amniotic and chorionic membranes, to these effects is still unknown. The aim of present study was comparison of the antibacterial effects between amniotic and chorionic membranes.<br /> Methods: Chorioamniotic layer was removed from placenta belonging to 43 healthy mothers whose infants were delivered by caesarean section. Their amniotic and chorionic fetal tissues were manually peeled in sterile conditions. The antibacterial effects of all membrane samples were evaluated on 8 standard strains of bacterial collection using disk diffusion method on bacteriologic media. Results of bacterial growth inhibition in the presence of amniotic or chorionic membranes were measured and recorded as median&plusmn;IQR. For data analysis and statistical comparison of samples, Kruskal-Wallis and Mann-Whitney U-test were applied using SPSS (v. 18).<br /> Results: Amniotic and chorionic membranes significantly showed different level of growth inhibitory effects on 8 bacterial strains including seven pathogens: <em>E. coli</em>, <em>Bacillus cereus</em>, <em>Klebsiella pneumonia</em>, <em>Streptococcus pyogenes</em>, <em>Pseudomonas aeruginosa</em>, <em>Staphylococcus aureus</em>, <em>Shigella flexneri</em> and one probiotic: <em>Lactobacillus plantarum</em> (p=0.018 and p&lt;0.001, respectively). The number of bacterial growth inhibition zones around chorionic membranes was more than of what found around amniotic membranes.<br /> Conclusion: The superiority of antibacterial effects of the chorionic membrane compared with the amniotic membrane can represent the key role of maternal part in placenta in protecting the fetus against possible infections. The antimicrobial effect of amniotic and chorionic membranes is significantly different on various bacteria.</p> https://www.jri.ir/article/701 Vitrification Increased Vacuolization of Human Spermatozoa: Application of MSOME Technology <p>Background: Sperm vitrification is a technique of ice and cryoprotectant free cryopreservation by direct plunging of sperm suspension into liquid nitrogen (LN2). The aim of this study was to investigate the influence of cryoprotectant free-vitrification on human sperm fine structure by MSOME technology and the fertility potential by zona binding assay (ZBA).<br /> Methods: 20 normo-ejaculates were prepared by swim up technique, and supernatants were divided into two parts of fresh and vitrified groups. For vitrification, sperm was dropped into LN2. Sperm motility, morphology, viability and MSOME were evaluated for each sample. In MSOM morphologically normal sperm (class 1), &le;2 small vacuoles (class 2), and one large vacuole or &gt;2 small vacuoles (class 3) were evaluated. Also, fertility potential was evaluated by zona binding assay. Data was analyzed using paired t-test or Willcoxon&rsquo;s test and p-value &lt;0.05 was considered significant.<br /> Results: Vitrification significantly reduced both progressive motility, viability and morphology. Also, normal morphology of spermatozoa decreased significantly after vitrification. In MSOME evaluation, normal motile spermatozoa (Class 1) decreased from 23.00&plusmn;12.44 to 16.00.56&plusmn;10.79 after vitrification (p=0.008). Although spermatozoa classes 2 and 3 were increased, the difference was not significant. Moreover, fertility potential of motile spermatozoa was reduced after vitrification (9.0&plusmn;13.87 <em>vs.</em> 13.40&plusmn;22.73; p=0.07).<br /> Conclusion: Vitrification increased the rate of vacuolization in motile sperm head. Therefore, MSOME technology is recommended for assessment of sperm fine morphology in ICSI program used cryopreserved spermatozoa.</p> https://www.jri.ir/article/702 Should Subclinical Hypothyroidism Be an Exclusion Criterion for the Diagnosis of Polycystic Ovary Syndrome? <p>Background: The purpose of the study was to examine whether patients with subclinical hypothyroidism (SCH) should be excluded before making a diagnosis of polycystic ovary syndrome (PCOS).<br /> Methods: Seven hundred sixteen patients, 462 with true PCOS, 31 with PCOS-SCH, and 223 normal cycling women were enrolled. Clinical, metabolic, and hormonal parameters among the groups were investigated. Continuous variables were compared by one-way analysis of variance. Proportions were compared using Z test. Fisher test was used to compare categorical variables. Simple correlation was performed using Spearman&rsquo;s coefficient. Correlation between thyroid stimulating hormone (TSH) and dependent variables were performed using backward multiple regression. The significance level was set at 0.05.<br /> Results: True polycystic ovary and polycystic ovary with subclinical hypothyroidism patients presented similar anthropometrical parameters. C-peptide was higher in polycystic ovary patients than in the other groups (p=0.014). Prevalence of glucose intolerance (p=0.186) and insulin resistance (p=0.293) was not statistically different in polycystic ovary and polycystic ovary with subclinical hypothyroidism. TSH levels showed positive correlation with lean body mass (p=0.032), total cholesterol (p=0.046, insulin (p=0.048) and prolactin (p=0.047). Backward multiple regression model retained TC, insulin, and PRL as predictors of TSH levels (p=0.011).<br /> Conclusion: Anthropometric parameters and ovary morphology were similar in both PCOS and PCOS-with-SCH patients. Regarding hormones, only C-peptide was higher in PCOS group. TSH correlated with total cholesterol, insulin, and prolactin. Before PCOS diagnosis, the exclusion criterion thyroid dysfunction should be standardized and subclinical hypothyroidism should not exclude a diagnosis of PCOS.</p> https://www.jri.ir/article/703 The Window of Implantation Is Closed by Estrogen via Insulin-Like Growth Factor 1 Pathway <p>Background: The opening and closing of the implantation window is important for successful pregnancy in eutherians. The recent study demonstrated that the window of uterine receptivity was prepared by the sole action of progesterone in mice, but the mechanism to close the window remained to be elucidated.<br /> Methods: The pregnant mice were ovariectomized on the evening on the third day of pregnancy with a single injection of medroxyprogesterone acetate to induce delayed implantation (DI). Several treatments were applied to DI mice. The uterine receptivity after treatment was assessed by examining cell proliferation in the uterine luminal epithelium (LE). The gene expressions in the endometrium were investigated by RNA-seq. The p&lt;0.05 was considered significant.<br /> Results: Cell proliferation in the LE ceased only when the window of implantation was open. Estrogen (E2) stimulated cell proliferation in the LE rendered the uterus refractory. The high throughput gene expression analysis by RNA-Seq showed that the insulin-like growth factor 1 (IGF1) pathway was the candidate to close the implantation window under E2. <em>In vivo</em> administration of IGF1 to delayed implantation mice resulted in proliferation in the LE cells.<br /> Conclusion: This study demonstrated that the window of uterine receptivity was closed by E2, which was mediated by the IGF1 pathway.</p> https://www.jri.ir/article/705 The Importance of Fertility Preservation Counseling in Patients with Gynecologic Cancer https://www.jri.ir/article/708 Maternal and Fetal Tuberous Sclerosis: Do We Know Enough as an Obstetrician? <p>Background: Tuberous sclerosis, also known as tuberous sclerosis complex (TSC), is a rare genetic condition that mainly causes hamartomas to develop in different parts of the body. TSC, an autosomal dominant trait with variable penetrance, can adversely affect maternal and fetal outcome.<br /> Case Presentation: In this paper, a case of maternal and fetal tuberous sclerosis having fetal cardiac rhabdomyoma detected in utero at 26 weeks was reported who&nbsp; subsequently had fetal demise at 31 weeks.<br /> Conclusion: Tuberous sclerosis is a rare genetic condition that mainly causes development of hamartomas. In tuberous sclerosis, a cardiac rhabdomyoma is the only sign that can be detected prenatally. In maternal tuberous sclerosis, fetal ECHO is advisable after 24 weeks. A pregnancy complicated by maternal or fetal tuberous sclerosis deserves careful observation and the fetus should undergo prenatal fetal Doppler echocardiography and if possible magnetic resonance imaging for evaluation of other fetal structures including brain and renal parenchyma, so that parents can be counseled regarding its future prognostic implications. Tuberous sclerosis can lead to poor fetal outcome including intrauterine fetal death; hence regular antenatal follow up is required. Genetic counseling is recommended for couples who have a family history of tuberous sclerosis and who want to have children. Prenatal diagnosis is available for families with a known gene mutation or history of this condition.</p> https://www.jri.ir/article/710 Poor Quality Embryos Hamper the Development of High Quality Ones https://www.jri.ir/article/711