Updated project metadata. Extracellular vesicles (EVs) participate in cell-stroma crosstalk within tumor microenvironment (TME) and Fb are key elements that contribute to tumor promotion. In thyroid cancer, the most prevalent endocrine malignancy, the relevance of Fb in the TME and EVs is beginning to be deciphered. Using an in vitro model of tumor-stroma crosstalk, we performed the comparative proteomic analysis of EVs secreted in tumoral- and non-tumoral milieu to describe EV-functionality and examine their possible role in thyroid tumor progression-related events. Tumor (TPC-1, 8505c) or non-tumor (NThyOri) thyroid cells were co-cultured with human fibroblasts (Fb). EVs, obtained by ultracentrifugation of conditioned media (CMs), were characterized by mass-spectrometry.