Toxoplasma gondii, the causative agent of toxoplasmosis, infects roughly one-third of the global population, highlighting its remarkable adaptability and pathogenicity across a wide range of warm-blooded animals. A key aspect of T. gondii’s ability to interact with its hosts involves the secretion of extracellular vesicles (EVs), which play a crucial role in mediating host-parasite interactions. These EVs carry a variety of molecular cargoes, including proteins, lipids, and RNA, which can alter host cellular responses and promote parasite survival. This project focuses on understanding how T. gondii adapts its EVs to the host environment. To explore this, we analyzed the EV cargo of T. gondii cultured in four different host cell lines representing distinct species. Each cell type provides a unique environment that mimics the conditions encountered by T. gondii during infection. By comparing the EV compositions across these cell lines, we aim to identify the diverse adaptive strategies that T. gondii employs through its EVs. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we characterized the EVs and conducted a comprehensive quantitative analysis of their proteomic cargo. Our findings revealed notable differences in the abundance and composition of EVs produced by tachyzoites cultivated in distinct host environments, underscoring the significant influence of host cell type on the makeup of T. gondii EVs.