The long-term adaptations to aerobic exercise training (AET) are diverse, multi-systemic, and include major health benefits and prolonged lifespan. Acute contractile activity stimulates the release of a myriad of bioactive factors, including cytokines, adipokines, and myokines (termed exercise factors or ‘exerkines’), which are postulated to drive training adaptations by exerting autocrine, paracrine, and endocrine effects. Recently, extracellular vesicles (EVs) have been proposed to act as carriers of exerkines, thereby ‘mediating’ cell-to-cell communication and inter-organ crosstalk. In vitro and in vivo studies have suggested that exercise augments the biogenesis and release of several EV sub-populations, including exosomes and microvesicles (MVs), and that they may be associated with the simultaneous release of classical myokines and/or cytokines (such as IL-6, IL-1, and TNFα). However, several inherent challenges to blood preparation procedures, EV isolation protocols, and classification of vesicle subpopulations, have delayed the advancement of our current understanding of exercise effects on the secretome. Independent variables, such as training status, gender, age, and disease also modulate EVs and exerkines (e.g., biogenesis, composition, secretion, and/or cargo), and must be considered when interpreting the growing body of knowledge. Because AET unequivocally decelerates biological aging, reduces all-cause mortality risk, and extends lifespan, academic and commercial interests are rapidly expanding across the globe, which necessitates methodological standardization and consistency between studies in order to generate high-quality, reproducible data.