Dosil, Sara G.Sánchez Madrid, FranciscoFernández Messina, Lola María2025-06-042025-06-042025-04Dosil SG, Sánchez‐Madrid F, Fernández‐Messina L. Isolation and Characterization of Extracellular Vesicles from Natural Killer Cells. Current Protocols 2025;5:e70125. https://doi.org/10.1002/cpz1.70125.2691-129910.1002/cpz1.70125https://hdl.handle.net/20.500.14352/120933Funding: This study was funded by the AECC (grant no. PRYCO223003PEIN), the Spanish Ministry of Science and Innovation (grant no. PID2023\u2010149541OB\u2010I00), and La Caixa Banking Foundation (grant no. HR23\u201000837).Natural killer (NK) cells are critical components of the immune system, recognized for their ability to identify and eliminate target cells undergoing various forms of stress. Beyond their direct cytotoxic activity, NK cells secrete extracellular vesicles (EVs), which serve as mediators of intercellular communication. These vesicles play an essential role in modulating immune responses and influencing tumoral and post-infection microenvironments. In this study, we present a detailed methodology for the isolation and expansion of human primary NK cells, the accumulation of their EVs, and the subsequent procedures for their isolation and characterization.engjournal articlehttps://doi.org/10.1002/cpz1.70125https://currentprotocols.onlinelibrary.wiley.com/doi/abs/10.1002/cpz1.70125metadata only access576.3612.017.4ExosomesExtracellular vesiclesEVsLymphocytesNatural killer cellsNK cellsPeripheral blood mononuclear cellsBiología celular (Biología)Inmunología2407 Biología Celular2412 Inmunología