Follicular fluid exosomes were isolated and characterized according to a previously published protocol. 17 The follicular fluid was gradually melted on ice and diluted by adding 20 mL PBS (pH 7.4) (Thermo Fisher Scientific). The follicular fluid was isolated by centrifugation at 2500 g for 30 minutes, and the supernatant was then transferred to a new centrifugal tube and centrifuged at 12 000 g for 5 minutes to eliminate large particles. After centrifugation, we filtered the supernatant using a 0.22 μm filter. Finally, the filtered samples were transferred to ultracentrifuge tubes for centrifugation at 120 000 g and 4°C for 4 hours in an ultracentrifuge (Beckman Coulter). Thereafter, exosome pellets were re‐suspended in RIPAlysate (Thermo Fisher Scientific) for western blot analysis or in PBS for nanoparticle tracking analysis (NTA). Then, the exosomes were collected for further treatment. Through the dynamic light scattering method, the particle size distribution of exosomes was evaluated. We entrusted the NTA analysis to Shanghai XP Biomed Ltd.

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