Clinical predictors of human blastocyst formation and pregnancy after extended embryo culture and transfer.

OBJECTIVE: To determine the clinical factors associated with blastocyst development and pregnancy. DESIGN: Evaluation of a prospectively collected IVF database. SETTING: An academic IVF practice. PATIENT(S): Couples (n = 529) undergoing their first IVF cycle who qualified for extended embryo culture (more than six zygotes) over the past 5 years. INTERVENTION(S): Seven or eight zygotes were cultured for 5 days, assessed for quality, and then transferred with excess zygotes cryopreserved at the pronuclear stage. MAIN OUTCOME MEASURE(S): Clinical predictors of blastocyst development and pregnancy. RESULT(S): The mean blastocyst development rate was 49.8%, with a median number of total and good-quality blastocysts of 4 and 2, respectively. Clinical factors that were significantly associated with good-quality blastocyst formation were younger female age, increased parity, standard insemination, and lower doses of gonadotropins. Clinical factors that were significantly associated with successful pregnancy were younger female age, higher antral follicle counts, greater numbers of total and good-/excellent-quality blastocysts, and absence of male factor infertility. CONCLUSION(S): Several clinical factors are associated with the development of good-quality blastocysts after extended embryo culture and successful pregnancy outcome. These patient and cycle characteristics may be very useful in selecting the best candidates for extended embryo culture and single blastocyst transfers, thus optimizing outcomes while reducing the risks associated with multiple pregnancies.

Effects of exogenous testosterone supplementation in gonadotrophin stimulated cycles.

BACKGROUND: Various experiments suggest that ovarian follicular recruitment and growth may be increased by testosterone priming. Our aim was to determine the effects of exogenous testosterone supplementation in older women on ovarian folliculogenesis and steroidogenesis. METHODS: A prospective randomized double-blind placebo-controlled crossover study was carried out. Twelve regularly menstruating non-obese women aged 38-45 years received a 12-day course of transdermal testosterone (2.5 mg per patch) or placebo patch, followed by 7 days of gonadotrophin stimulation. After at least a 1 month washout period, subjects underwent the same protocol using the opposite treatment. The main outcomes were follicular development (ultrasound measures) and hormone levels. RESULTS: Following gonadotrophin stimulation, there were no differences in average number of follicles over 10 mm diameter in cycles pre-treated with testosterone versus placebo [2.10 (95% confidence interval (CI) 1.11, 3.22) versus 2.08 (95% CI 1.03, 3.14), P = 0.55]. No crossover, period (first or second test) or sequence (order of treatment) effects were noted. As expected, total and free testosterone levels were increased following testosterone treatment (312.7 +/- 122.4 versus 12.3 +/- 4.5 ng/dl and 45.5+/- 16.7 versus 1.4 +/- 0.5 ng/dl, respectively, P < 0.001) but no differences in free or total testosterone were noted by period. LH, FSH, estradiol and antral follicle counts before gonadotrophin stimulation were not altered by testosterone pretreatment or by period. CONCLUSIONS: Despite increased testosterone levels, a short course of androgens had no significant effect on the number of follicles over 10 mm during stimulation with FSH in women of late reproductive age.

An in vitro model to study the pathogenesis of the early endometriosis lesion.

OBJECTIVE: To determine if whole fragments of endometrium can adhere to peritoneum with intact mesothelium. DESIGN: Tissue culture and immunohistochemical study. SETTING: University Medical Center. PATIENTS: Reproductive-age women undergoing surgery for benign conditions. INTERVENTIONS: Whole explants of human peritoneum from the anterior abdominal wall and the posterior surface of the uterus were cultured with whole fragments of mechanically dispersed endometrium. MAIN OUTCOME MEASURES: Adhesion of endometrial fragments to the surface of the peritoneum was evaluated. Adherent fragments of endometrium were identified using the dissecting microscope and by performing serial sections of the peritoneum explants for light and confocal laser-scanning microscopy. Immunohistochemical staining of the mesothelium with antibodies to cytokeratin and vimentin was used to ensure an intact layer of mesothelium beneath the endometrial implants. Transmission electron microscopy was also used to evaluate the adhesion of endometrium to the mesothelium. RESULTS: Endometrium was identified attached to the surface of the peritoneum. After 18-24 hours of culture, the majority of implants did not have identifiable mesothelium beneath them, but most had intact mesothelium running up to the point of attachment. Approximately 10% of the endometrial implants had intact mesothelium at the site of attachment. After 1 hour of culture, both endometrial stromal and epithelial cells were attached to intact mesothelium in nearly all cases. Early transmesothelial invasion involves endometrial stromal cells. CONCLUSIONS: Endometrial stromal and epithelial cells can attach to the intact mesothelial surface of the peritoneum. Endometrial stromal cell invasion through the mesothelium occurs in less than 18-24 hours.