Meso–macroporous hydrogel for direct litre-scale isolation of extracellular vesicles

Abstract Extracellular vesicles are cell-originated lipid bilayer membrane vesicles that play vital roles in cell-to-cell communications. While extracellular vesicles hold substantial biomedical potential, conventional methodologies for isolating extracellular vesicles require elaborate preprocessing and, therefore, remain labour intensive and limited by throughput. To overcome these challenges, we present a facile fabrication route for generating a meso–macroporous hydrogel matrix with pores of ~400 nm for customizable extracellular vesicle isolation. By combining surface charge-selective capture of extracellular vesicles within the hydrogel matrix and their recovery by high ionic strength, we report direct extracellular vesicle isolation with a throughput range from microlitre to litre scales, without preprocessing, for various biofluids, including whole blood, plasma, ascites, saliva, urine, bovine milk and cell culture media. Furthermore, we demonstrate that the meso–macroporous hydrogel also serves as a solid-phase matrix for preserving extracellular vesicles for on-demand downstream analyses, making it applicable for therapeutics, cosmeceuticals and disease diagnostics.