The neonatal mammalian heart is able to regenerate after injury by inducing cardiomyocyte proliferation. However, this regenerative capacity is virtually lost in the adult mammalian heart. Extracellular vesicles (EVs) have been shown to play an important cardioprotective role in heart repair. Here, we performed proteomic analysis of EVs from neonatal mouse heart tissues (Neo-EVs), EVs regenerated from neonatal heart tissues after apicoectomy (AR-Neo-EVs), and EVs from adult mouse hearts (Adu-EVs), to compare the differential changes in proteins among them.