fig1

Figure 1. Schematic drawing for extracellular vesicle isolation procedure by differential ultracentrifugation. For extracellular vesicle isolation, the supernatant is collected from cells cultured in dishes for 48 h at 37 °C. In the first step, dead cells and cell debris are spun down from the supernatant at 2000 × g for 20 min at 4 °C. The supernatant collected is spun at 9000 rpm (10,000 × g) for 35 min at 4 °C. The pellet obtained corresponds to the LEVs and contaminating proteins. The supernatant collected without disturbing the 10,000 × g pellet is spun at 100,000 × g for 70 min at 4 °C. The pellet corresponding to sEVs and contaminating proteins are washed in PBS followed by a second spin at 100,000 × g for 70 min at 4 °C. The final sEV pellet is resuspended in PBS and transferred to Eppendorf tubes. The collected EVs can be biochemically and functionally characterized. sEVs: small extracellular vesicles; LEVs: large extracellular vesicles