Figure 1.

Flowchart showing methods for the isolation of extracellular vesicles (EVs) from milk and subsequent exosome enrichment. EVs were isolated from human and bovine milk by differential ultracentrifugation. Unpasteurised milk was centrifuged at 3000 rcf followed by 0.25M EDTA (1 : 1; v/v) treatment to remove excess casein and supernatants (S/N) subsequently centrifuged at 12,000, 30,000, 70,000, and 100,000 rcf, respectively. The pellet obtained after the sequential centrifugation process contains EVs. After reconstitution in PBS, the EV suspension was used for exosome enrichment. 600 μl of EV suspension was introduced on top of a SEC column (qEV column) and processed via SEC to obtain 16 fractions as described in the flowchart.