Cells from organisms across all kingdoms of life continuously release a diverse repertoire of extracellular vesicles (EVs) into their extracellular environment as an elegant strategy for both, cellular homeostasis and communication with other cells. Through different biogenesis routes within the donor cell, nanosized vesicles are generated either from endomembranes or the plasma membrane, and loaded and decorated with macromolecular cargo in a controlled manner through molecular sorting machineries. Since they can affect a recipient cell in the same tissue, distant organs or even other organisms, EVs have been increasingly recognized as essential mediators orchestrating intercellular communication in health and disease. In the last 15 years, research on the fundamental biology of EVs as well as their potential for biomedical applications has been greatly intensified. Time to present new advances on EV biogenesis, their intercellular communication competencies as well as technical and biomedical applications in a special thematic series of Cell Communication and Signaling.