A visualized and quercetin-optimized three-dimensional culture model of mouse ovaries derived from fetal gonads.

Abstract

The in vitro culture of ovarian tissue is emerging as a popular technology to study female reproductive medicine. However, standard in vitro culture conditions usually increase the level of reactive oxygen species (ROS), hindering ovarian development. Here, we establish an in vitro visualized mouse ovarian explant 3D culture model with the GFP-BVSC reporter system and obtain the early follicle pool from fetal female gonads. This model recapitulates in vivo ovarian characteristics and allows non-invasive monitoring of ovarian development. Importantly, supplementation with quercetin, a plant-derived natural antioxidant, increases the tissue area and total follicle count in cultured ovaries by protecting mitochondria and reducing ROS, thus more closely mimicking in vivo growth conditions. Finally, this visualized and optimized ovarian explant culture platform has been proven to be effective in modelling female ovarian diseases, such as the fetal reproductive aberrations of female offspring affected by gestational diabetes mellitus (GDM). Overall, our work extends the understanding of ovarian biology and creates an efficient and simplified platform for the morphological monitoring of ovarian development, as well as for drug screening and the clinical treatment of ovarian hypofunction.