An engineered ovarian somatic organoid model exhibits age-dependent changes in cellular organization and function

Ovarian somatic cells are essential for tissue function, but there are no ex vivo models which maintain the cellular heterogeneity, in addition to the cell-cell and cell-matrix interactions of this compartment. We engineered a novel ovarian somatic organoid model in which a stroma-enriched fraction of primary mouse ovarian somatic cells was cultured in scaffold-free agarose micromolds. We sought to use this ovarian somatic organoid model to investigate mechanisms of ovarian aging and performed single cell RNA sequencing using primary ovarian somatic cells isolated from reproductively young and old mice, as well as organoids generated from young and old mice at days 1 and 6 of culture to assess age-dependent changes to cellular composition. Primary ovarian somatic cells isolated from reproductively young (6-12 weeks) or old (10-14 months) mice were lifted from 2D culture overnight or dissociated from organoids at day 1 or day 6 of culture and analyzed by scRNAseq