Data and code availability •The full proteomics mass spectrometry data were uploaded into the PRoteomics IDEntifications Database (PRIDE) and are publicly available as of the date of publication. (65) EV isolation methods and study logistics have been uploaded to the EV-Track knowledgebase (EV-TRACK ID: EV230971). (66) Cell viability, cell permeability, NTA, and proteomics data were also uploaded to the UNC Center for Environmental Medicine, Asthma and Lung Biology (UNC-CEMALB)-Dataverse. (67) The PRIDE project accession number, EV-TRACK ID, and accession link to UNC-CEMALB-Dataverse are listed in the key resources table. •Script used to analyze NTA and proteomics data were organized via the UNC-CEMALB Github website and is publicly available as of the date of publication. (68) •Any additional information required to reanalyze the data reported in this paper is available from the lead contact upon request. This dataset is associated with the following publication: Vitucci, E., C. Carberry, A. Payton, L. Herring, A. Mordant, S. McCullough, and J. Rager. Characterizing the Extracellular Vesicle Proteomic Landscape of the Human Airway using In vitro Organotypic Multi-Cellular Models. iScience. Elsevier B.V., Amsterdam, NETHERLANDS, 26(11): 108162, (2023).