Adipose tissue-derived extracellular vesicles (ATEVs) have garnered attention for their roles in intercellular communication and regulation. This study presents the first investigation of equine adipose tissue, with ATEV being extracted concurrently during the isolation of adipose-derived mesenchymal stem cells (ADSCs). Through CCK-8 cell proliferation assays and scratch migration assays, a significant promotional effect of ATEV on ADSCs was observed, which not only accelerated the proliferation rate of the stem cells but also enhanced their migratory capacity. The application of Trypan Blue exclusion and live-dead cell staining further demonstrated the positive effects of ATEV on maintaining the viability of ADSCs during ex vivo storage. These findings provide new insights into the potential applications of ATEV in stem cell therapy, tissue repair, and regenerative medicine.