Human ovarian granulosa cells and follicular fluid indices: the relationship to oocyte maturity and fertilization in vitro

The study investigates the correlation between oocyte maturity and fertilization and a variety of hormonal parameters in follicular fluid and ovarian granulosa cells. A methodology for purification of granulosa cells from contaminating blood cells is also established. A total of 63 follicular aspirates were collected at oocyte retrieval from 30 women superovulated using the long luteinizing hormone- releasing hormone (LHRH analogue)/human menopausal gonadotrophin regimen. Oestradiol, progesterone, testosterone and human chorionic gonadotrophin (HCG) were quantified in follicular fluid and granulosa cells were immunostained for human chorionic gonadotrophin. Immunopurification of granulosa cells from contaminating blood cells was performed. HCG in follicular fluid was significantly high in follicles yielding immature (grade 3) oocytes (P=0.002); there was no correlation with fertilization. Aspirates from follicles containing mature (grade 1) oocytes and oocytes that subsequently fertilized had significantly more granulosa cells immunobound to HCG (P < 0.001, P=0.02). Moreover, the immunomagnetic purification technique provided >98% pure population of granulosa cells. The data demonstrate that HCG in follicular fluid and on granulosa cells may help to predict oocyte maturity and fertilization. Furthermore, immunomagnetic beads provide a reliable procedure for the purification of ovarian granulosa cells.      

Association of abnormal vaginal flora with increased cervical tumour necrosis factor--alpha and interferon--gamma levels in idiopathic infertility

This study was conducted to evaluate the presence of bacterial vaginosis (BV) and its association with cervical tumour necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) levels in idiopathic infertility. Forty idiopathic infertile women and twenty fertile controls were recruited from the Department of Obstetrics and Gynaecology, El Shatby Maternity Hospital, Alexandria. High vaginal swabs were smeared and assessed by Gram staining for the presence of BV using Nugent's scoring system. Cervical mucus samples were collected at midcycle and assayed for IFN-gamma and TNF-alpha by enzyme linked immunosorbent assay (ELISA). Bacterial vaginosis was identified in 25% (10/40) of the infertile group and in 10% (2/20) of controls. The mean concentrations of TNF-alpha (232.2 +/- 51.6 pg/ml) and IFN-gamma (127 +/- 26.8 pg/ml) were significantly higher in the infertile group compared with controls (P < 0.001). Infertile women with BV showed significantly higher cervical levels of TNF-alpha (649 +/- 126.8 pg/ml) and IFN-gamma (350 +/- 59.2 pg/ml) than those with normal (62.2 +/- 8.1, 31.4 +/- 4.1 pg/ml respectively) and intermediate (252.5 +/- 21.4, 170 +/- 17.3 pg/ml respectively) vaginal flora (P < 0.001). A significant correlation was found between TNF-alpha and IFN-gamma concentrations in the idiopathic infertile group (r = 0.984, P < 0.001) as well as the fertile control group (r = 0.881, P < 0.001). In conclusion, BV is associated with elevated cervical mucus levels of TNF-alpha and IFN-gamma. The induction of these proinflammatory cytokines by an altered vaginal ecosystem may play a role in the etiology of idiopathic infertility. This may have potential applications in the diagnosis and treatment of female infertility.     

Human chorionic gonadotrophin and steroid concentrations in follicular fluid: the relationship to oocyte maturity and fertilization rates in stimulated and natural in-vitro fertilization cycles

The study investigates the relationship of follicular fluidsteroids and human chorionic gonadotrophin to oocyte maturityand fertilization rates in stimulated and natural cycles. Oestradiol,progesterone, testosterone and human chorionic gonadotrophinwere quantified in 129 samples of follicular fluid and the progesterone:oestradiol ratio calculated. Both stimulated cycles (short andlong luteinizing hormone-releasing hormone/human menopausalgonadotrophin regimens) and natural cycles were compared. Atotal of 60 women were studied, 20 in each group. In the naturalcycles, testosterone was significantly lower in follicles withintermediate oocytes (P = 0.015). Both oestradiol and testosteronewere significantly lower in stimulated cycles compared to naturalcycles (P = 0.032 and P = 0.034 respectively). In the ovarianstimulation cycles, the progesterone: oestradiol ratio was significantlyhigher when oocytes fertilized (P = 0.052). Moreover, in thestimulated cycles, oestradiol and human chorionic gonadotrophinwere significantly lower in the short protocol compared to thelong protocol. The data demonstrate that the hormonal milieuof the follicle is altered in downregulated stimulated cyclesto varying degrees, depending partially on the type of protocolused. Furthermore, the progesterone: oestradiol ratio, ratherthan individual hormone concentrations, may be a useful predictorof the fertilizing capacity of the oocytes.