Connecting the Dots: Milestones in the History of Extracellular Vesicle Research.

The field of extracellular vesicle (EV) research offers a compelling example of a biological concept refined through continuous methodological innovation. This review traces the historical trajectory of the discipline chronologically, beginning with early observations in haemostasis, from Malpighi's descriptions of blood clots and Chargaff and West's identification of a procoagulant sedimentable plasma fraction, to Wolf's "platelet dust," Crawford's microparticles characterised by electron microscopy, and the seminal work by Stahl and Johnstone demonstrating regulated vesicle biogenesis during reticulocyte maturation via multivesicular bodies. We highlight a pivotal conceptual shift, from viewing EVs as cellular debris to recognising them as regulated "communicasomes," catalysed by Raposo's discovery of antigen-presenting exosomes and subsequent evidence for EV-mediated transfer of functional receptors and nucleic acids, including the influential and sometimes debated model proposed by Ratajczak. By integrating findings from matrix vesicles, plant-derived vesicles, and diverse tissue contexts, we frame EV release as an evolutionarily conserved process with profound implications for immunity, regeneration, oncology, and cardiovascular pathology. A second central aim of this review is practical and methodological. We map how the expansion of biological claims has driven urgent standardisation efforts, notably through the establishment of the International Society for Extracellular Vesicles (ISEV) and the successive MISEV guidelines (2014, 2018, 2023). These are complemented by community resources such as EV-TRACK, MIFlowCyt-EV, and the databases ExoCarta and Vesiclepedia. We summarise core experimental choices across isolation and characterisation techniques, including ultracentrifugation, size exclusion chromatography, density gradients, flow cytometry, nanoparticle tracking analysis, and electron microscopy, while outlining persistent bottlenecks in purity, standardised nomenclature, and experimental reproducibility. Finally, we provide concise biographical sketches of key contributors and an overview of major EV-focused journals and ISEV meetings that anchor consensus-building and the translation of fundamental knowledge into clinical and industrial applications.