Protocol for sorting hepatocyte-derived extracellular vesicles from mouse liver tissues using immunomagnetic beads.

Here, we present a protocol for isolating total liver tissue-derived extracellular vesicles (LT-EVs) from mouse liver tissue and purifying hepatocyte-derived extracellular vesicles (Hep-EVs) using asialoglycoprotein receptor (ASGPR)-based immunomagnetic sorting. We describe steps for mouse liver perfusion, enzymatic digestion, differential centrifugation, ASGPR labeling, and magnetic-activated cell sorting (MACS)-based enrichment. We then detail procedures for extracellular vesicle (EV) quality control by flow cytometry, nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blotting. For complete details on the use and execution of this protocol, please refer to Ying et al.1.