Towards a systematic understanding of the diverse subtypes, biogenesis and functions of large extracellular vesicles.

Large extracellular vesicles (lEVs) are heterogeneous structures with sizes that exceed 200 nm. Unlike small extracellular vesicles, which originate from either the endosomal system or the plasma membrane, most lEVs stem directly from the plasma membrane, although the various lEV subtypes have distinct biogenesis pathways. Emerging findings have helped to update lEV classification and have provided insights into lEV biogenesis, cargo, physiological functions and relevance in disease. Classical lEVs include apoptotic bodies, microvesicles and large oncosomes, whereas detached migrasomes, exophers, mitophers, blebbisomes, large ageing neutrophil-derived vesicles were only identified later. The study of these diverse lEV subtypes has been challenging, but emerging technologies, including tools that enable multidimensional characterization, loss-of-function studies and mechanistic analyses, are likely to advance the field further, especially if they are appropriately integrated to ensure rigour and reproducibility.