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Objective: To evaluate the efficacy of a microfluidic culture system supplemented with follicular fluid meiosis-activating sterol (FF-MAS) on the maturation of immature oocytes in patients with polycystic ovarian syndrome (PCOS)., Methods: A total of 438 germinal vesicle oocytes from 163 PCOS patients were included. Oocytes were divided into five groups: (1) cultured in static drops without FF-MAS, (2) cultured in static drops with FF-MAS, (3) cultured in a microfluidic device without FF-MAS, (4) cultured in a microfluidic device with FF-MAS for the first 2 h, and (5) cultured in a microfluidic device with FF-MAS for 24 h. Maturation, fertilization, and embryo development rates were assessed, alongside gene expression analysis using quantitative real-time PCR. Ultrastructural analysis was performed using transmission electron microscopy (TEM)., Results: The maturation rates for oocytes in groups 1 to 5 were 36%, 48%, 42%, 64%, and 65%, respectively, with groups 4 and 5 showing significant increases (p < 0.05). Fertilization rates were higher in groups 4 and 5 (p < 0.05). High-quality embryo formation rates were significantly higher in groups 3, 4, and 5. Gene expression analysis indicated enhanced expression of BRCA1 and TP53 in the dynamic culture groups. Ultrastructural analysis showed improved mitochondrial integrity and cytoplasmic maturation in the dynamic groups., Conclusions: The dynamic microfluidic culture system with FF-MAS supplementation significantly enhances oocyte maturation and embryo quality in PCOS patients by closely mimicking the natural in vivo environment. This approach shows promise for optimizing in vitro oocyte maturation and improving fertility outcomes., (© 2025 Japan Society of Obstetrics and Gynecology.)