A comparison of polarized and non-polarized human endometrial monolayer culture systems on murine embryo development

BACKGROUND: Co-culture of embryos with various somatic cells has been suggested as a promising approach to improve embryo development. Despite numerous reports regarding the beneficial effects of epithelial cells from the female genital tract on embryo development in a co-culture system, little is known about the effect of these cells when being cultured under a polarized condition on embryo growth. Our study evaluated the effects of in vitro polarized cells on pre-embryo development. METHODS: Human endometrial tissue was obtained from uterine specimens excised at total hysterectomy performed for benign indications. Epithelial cells were promptly isolated and cultured either on extra-cellular matrix gel (ECM-Gel) coated millipore filter inserts (polarized) or plastic surfaces (non-polarized). The epithelial nature of the cells cultured on plastic was confirmed through immunohistochemistry, and polarization of cells cultured on ECM-Gel was evaluated by transmission electron microscopy (TEM). One or two-cell stage embryos of a superovulated NMRI mouse were then flushed and placed in culture with either polarized or non-polarized cells and medium alone. Development rates were determined for all embryos daily and statistically compared. At the end of the cultivation period, trophectoderm (TE) and inner cell mass (ICM) of expanded blastocysts from each group were examined microscopically. RESULTS: Endometrial epithelial cells cultured on ECM-Gel had a highly polarized columnar shape as opposed to the flattened shape of the cells cultured on a plastic surface. The two-cell embryos cultured on a polarized monolayer had a higher developmental rate than those from the non-polarized cells. There was no statistically significant difference; still, the blastocysts from the polarized monolayer, in comparison with the non-polarized group, had a significantly higher mean cell number. The development of one-cell embryos in the polarized and non-polarized groups showed no statistically significant difference. CONCLUSION: Polarized cells could improve in vitro embryo development from the two-cell stage more in terms of quality (increasing blastocyst cellularity) than in terms of developmental rate

Symptoms of natural menopause among Iranian women living in Tehran, Iran

Background: Menopause is a step of a woman’s life when hormonal changes cause menstruation to stop permanently. Menopausal symptoms can affect women's health and differ between different races and societies. Objective: The aim of this study was to survey the symptoms associated with menopause among Iranian women living in Tehran, Iran. Materials and Methods: In this cross sectional study which has been done between 2004 and 2005, women aged equal or more than 35 years old living in districts of Tehran were selected by multistage randomized cluster sampling. For each woman a questionnaire was completed. The data gained from each questionnaire was analyzed by using SPSS version 13. Results: At time of study, 2462 women were naturally menopause. The mean age of natural menopause onset was 47.71 (SE=0.11) years. In 52.9% of cases, the onset of menopause was sudden. The symptoms associated with menopause were night sweats (61.2%), joint and muscle pain (59.9%), hot flashes (53.1%), fatigue (45.6%), decreased libido (33.9%), insomnia (33.7%), weight gain (30.1%), forgetfulness (24.9%) and urinary symptoms (17.4%). Conclusion: This study showed that night sweats, joint and muscle pain and hot flashes are the most common symptoms associated with menopause

Effect of β-Mercaptoethanol with and without BSO (DL-Buthionine Sulfoximine) on Resumption of Meiosis, in vitro Maturation and Embryo Development of Immature Mouse Oocytes

Introduction: The purpose of this study was to evaluated the effect of β-mercaptoethanol on resumption of meiosis, in vitro maturation of immature mouse oocytes and resulting embryo development with and without BSO (DL-Buthionine sulfoximine). Material and Methods: Germinal vasicle (GV) were recovered from 6-8 weeks old NMRI ovaries and cultured in maturation medium in MEMα supplemented with 7.5IU/ml hCG, 100mIU/ml rhFSH, 5% FCS (control group) and adding 100μm β-mercaptoethanol (group 1) or with 5mM BSO + 100μm β-mercaptoethanol (group 2) for 24h. The matured oocytes then were fertilized and cultured for 5 days. Fertilization and development were accomplished in T6 medium. Results: The percentage of GV oocytes reaching to metaphase I (or undergo GVBD) were 78.5%, 85%, 86% in control group, group 1 and group 2 respectively, that no significant difference was detected between groups. The proportion of oocytes that progressed to the metaphase II (MII) stage was minimum within 5mM BSO group (group 2) and maximum within β-mercaptoetanol group (group 1) with significant difference comparing with control and each other (P≤0.05). The percentage of embryos reaching to morula stage within β-mercaptoetanol group was significantly higher than the control group (5% and 12.2% respectively). None of oocytes treated with BSO could pass the 8 cell stage. Conclusion: β-mercaptoetanol enhances IVM and improves embryo development. While adding BSO into the maturation medium even with β-mercaptoetanol decreases maturation and declines the embryo development