EV-Tracer enables lineage-resolved detection and molecular profiling of extracellular vesicle-associated signals in cancer–fibroblast co-culture
Year: 2026
Type: preprint
Abstract: Extracellular vesicles (EVs) facilitate intercellular communication by transferring diverse bioactive molecules from donor to recipient cells. However, EVs released by distinct cellular lineages become difficult to distinguish when mixed in multicellular experimental models, limiting the analysis of how cell–cell interactions affect EV-associated molecular profiles. To address this, EV-Tracer, a CD63-based dual-fluorescence tracing and capture system for detecting, isolating, and profiling lineage-associated EV fractions, is developed. Achilles or mScarlet is inserted into the small extracellular loop of CD63, enabling tracer-specific EV detection by digital counting, antibody-based isolation, and live-cell visualization. Exploratory EV RNA sequencing suggests that physical cell–cell contact is associated with distinct EV RNA profiles, including interferon-related signals, which are supported by targeted cellular and EV-associated RNA analyses. EV-Tracer provides a practical framework for investigating lineage-associated EV dynamics and molecular signals in mixed-cell systems.
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7204224715