Activation of multiple receptors stimulates extracellular vesicle release from trophoblast cells

Reports of the stimulated release of extracellular vesicles (EVs) are few, and the mechanisms incompletely understood. To our knowledge, the possibility that the activation of any one of the multitudes of G‐protein‐coupled receptors (GPCRs) expressed by a single cell‐type might increase EV release has not been explored. Recently, we identified the expression of cholecystokinin (CCK), gastrin, gastrin/cholecystokinin types A and/or B receptors (CCKAR and/or –BR), and the bitter taste receptor, TAS2R14 in the human and mouse placenta. specifically, trophoblast. These GPCR(s) were also expressed in four different human trophoblast cell lines. The current objective was to employ two of these cell lines—JAR choriocarcinoma cells and HTR‐8/SVneo cells derived from first‐trimester human villous trophoblast—to investigate whether CCK, TAS2R14 agonists, and other GPCR ligands would each augment EV release. EVs were isolated from the cell‐culture medium by filtration and ultracentrifugation. The preparations were enriched in small EVs (