Endocrine Abstracts

JOINT3582

Introduction: Infertility is an issue of increasing concern with an estimated 1 in 6 adults affected during their lifespan. This highlights the need for more comprehensive insight about the biological events that can affect the female fertility potential during a lifetime, and the importance of a more detailed understanding of ovarian physiology. Thus, this project aims to establish a foetal and a prepubertal mouse ovarian ex vivo culture model. These tissue culture models will be an essential tool to study the effects of selected manipulations on ovarian development, and early folliculogenesis.

Methods: Three ex vivo culture approaches were tested using ovaries from foetal and prepubertal mice including culture; on agarose gel fragments, in hanging drops, and on porous membranes. Ovaries were cultured for 48 hours, and 7 days. After the culture period, the three methods were evaluated, and overall tissue preservation was compared. Tissue morphology was evaluated using Periodic Acid-Schiff staining, while proliferation and apoptosis was investigated using BrdU incorporation and cleaved PARP staining, respectively. Subsequently, the established ovarian cultures will used to study effects of selected treatment given through the culture media. Analysis will include quantification of germ cells (Oct4-positive cells) and follicles (from PAS sections) and immunohistochemical stainings for cell lineage markers such as, granulosa cells (AMH, CYP19A1), and theca cells (CYP11A1, CYP17A1). +.

Results: The analysis of the three culture methods and culture lengths are currently ongoing. The optimal culture method will be determined based on preservation of morphology, minimal apoptosis, and sustained cell proliferation.

Conclusion: Preliminary findings suggest that ex vivo culture of foetal and prepubertal mouse ovarian tissue is feasible with the tested culture approaches. Ongoing analysis will identify the most appropriate culture approach for future studies examining the effects of manipulating various signaling pathways involved in the regulation of ovarian development and function.