This study demonstrates that in HeLa cells, small ectosomes (EVs budding directly from the plasma membrane) are more prominent than exosomes (EVs formed within endosomes). They identify new protein markers to distinguish these two EV populations and show that the endosomal pH affects exosome secretion but not ectosome secretion, confirming their different origins.

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This study uses innovative live-cell imaging and proteomics to reveal novel insights into the biogenesis and secretion of exosomes and ectosomes. The identification of specific markers for these different sEV subpopulations is a significant advance in the field, with potential implications for understanding their distinct roles in intercellular communication. While the study has some limitations, the methodology is strong and the findings are compelling.