This study investigated the impact of exosomes, released by ovarian cells in Sebastes schlegelii, on the vitality of stored sperm from a novel perspective. Comprehensive identification and analysis of the contents of ovarian fluid exosomes during the early and late stages of sperm storage (Sst_E and Sst_L) were performed using proteomics and small RNA sequencing. This study aimed to elucidate the interactive effects between exosomes, the ovary, and sperm under two distinct sperm storage states.