Mitochondrial dysfunction in granulosa cells is associated with impaired proliferation and angiogenic support in women with polycystic ovarian syndrome and elevated AMH

This study aimed to investigate the relationship between mitochondrial function and angiogenic capacity in granulosa cells derived from patients with PCOS. Specifically, we evaluated cellular proliferation, mitochondrial activity, and paracrine angiogenic support, and further examined whether these alterations vary according to follicular stage. Granulosa cells (GCs) were isolated from women undergoing IVF. PCOS was diagnosed according to the Rotterdam criteria. The final study cohort comprised a normal group with serum AMH levels of 2–5 ng/mL ( n = 12) and a PCOS group fulfilling the Rotterdam criteria with elevated AMH levels (> 5 ng/mL; n = 11). Cells were further classified by follicular diameter into large (> 14 mm) and small (< 14 mm) follicle subgroups. GCs were cultured under standardized conditions, and cellular morphology, proliferation, mitochondrial function, intracellular ATP levels, paracrine angiogenic capacity (HUVEC tube formation assay), and expression of angiogenesis-related cytokines were assessed. PCOS-derived granulosa cells (GCs) exhibited abnormal morphology, increased cell size, and reduced proliferative capacity compared with normal controls. Mitochondrial function and intracellular ATP levels were significantly decreased in PCOS GCs. Conditioned media from PCOS GCs impaired endothelial tube formation, indicating reduced paracrine angiogenic capacity, with a more pronounced effect observed in GCs derived from larger follicles. At the molecular level, key pro-angiogenic chemokines, including CXCL6, IL8, and MCP1, were consistently downregulated in PCOS GCs, whereas VEGF-A showed a less consistent pattern. PCOS granulosa cells exhibit coordinated impairments in cellular growth, mitochondrial function, and angiogenic support. These findings support a functional interplay between metabolic dysfunction and disrupted chemokine-mediated angiogenic signaling, providing a mechanistic framework for follicular impairment in PCOS and highlighting potential targets for improving reproductive outcomes.