Phthalates and bisphenol do not accumulate in human follicular fluid.

OBJECTIVE: To determine if phthalates and bisphenol A accumulate in human follicular fluid after brief exposure to medical plastics during an IVF cycle STUDY DESIGN: Prospective collection of follicular fluid from five infertile women undergoing oocyte retrieval at a University IVF laboratory and analysis of Phthalate & Bisphenol A levels. RESULTS: All phthalate levels were detected at levels less than 15 ng/mL and Bisphenol A levels were undetectable in all five samples. The concentrations of phthalates are 200-1000 fold less than the minimum levels reported to cause reproductive toxicity in vitro to cumulus-oocyte complexes of laboratory animals. CONCLUSIONS: In reproductive age women undergoing infertility treatments there is little transfer or accumulation of phthalates, phthalate metabolites or bisphenol A into the microenvironment of the human preovulatory oocyte and the levels are not clinically significant. Further investigation of phthalate and bisphenol A accumulation in vivo in human follicular fluid may not be productive.

In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary.

PURPOSE: create a 3-Dimensional artificial human ovary to mature human oocytes. METHODS: theca and granulosa cells were isolated from antral follicles of reproductive-aged women, seeded into micro-molded gels and self-assembled into complex 3D microtissues. Immunohistochemistry and live-dead staining confirmed theca cell identity and cellular viability at one week respectively. Placement of granulosa cell spheroids or cumulus-oocyte complexes into theca cell honeycomb openings resulted in creation of an artificial human ovary. Oocytes from this construct were assessed for polar body extrusion. RESULTS: theca and granulosa cells self-assembled into complex microtissues, remaining viable for one week. At 72 h after artificial human ovary construction, theca cells completely surrounded the granulosa spheroids or COCs without stromal invasion or disruption. Polar body extrusion occurred in one of three COCs assessed. CONCLUSIONS: an artifical human ovary can be created with self-assembled human theca and granulosa cell microtissues, and used for IVM and future oocyte toxicology studies.

Expression and localization of the novel and highly conserved gametocyte-specific factor 1 during oogenesis and spermatogenesis.

OBJECTIVE: To determine the onset of gametocyte-specific factor 1 (Gtsf1) expression in embryogenesis and its relation to Nobox; and to determine its localization during gonadal development and gametocyte maturation. DESIGN: Developmental animal study. SETTING: University reproductive biology laboratory. ANIMAL(S): Mice ranging in age from embryonic day 12.5 to 8 weeks. INTERVENTION(S): Polymerase chain reaction and quantitative polymerase chain reaction were performed to determine the onset of and relative messenger RNA expression. Western blot was performed to confirm protein expression and antibody specificity. In situ hybridization and immunohistochemistry were used determine localization of expression. MAIN OUTCOME MEASURE(S): Gtsf1 messenger RNA expression levels during embryogenesis through adulthood in wild-type mice and in newborn Nobox knockout mice; GTSF1 expression and localization in postnatal mice. RESULT(S): Gtsf1 functions downstream of Nobox and is highly expressed in embryonic male and female gonads, localizing to germ cells throughout development. GTSF1 expression is confined to the cytoplasm in all stages of postnatal oocyte maturation and to prespermatogonia during early postnatal testicular development. CONCLUSION(S): The expression pattern of Gtsf1 and its high conservation suggests that it may play an important role in germ cell development. Further characterization of Gtsf1 may elucidate mechanisms involved in premature ovarian failure.