Optimized Protocol for the Isolation of Extracellular Vesicles from the Parasitic Worm Schistosoma mansoni with Improved Purity, Concentration, and Yield

Figure 3

Iodixanol gradients are preferred over sucrose gradients for purification of adult worm and schistosomula EVs. EV-enriched UC pellets of adult worm EVs (a) or schistosomula EVs (b) were loaded at the bottom (bottom-up) or on top (top-down) of a small iodixanol density gradient and centrifuged for two hours. Western blot analysis of collected density fractions for the EV marker TSP-2 shows that both EV types reached equilibrium densities in the small gradients. (c) Adult worm EVs were loaded at the bottom or top of small sucrose density gradients. Shown are western blot detections of TSP-2 in fractions collected from two and 13.5 hours centrifuged sucrose density gradients. (d) Top-down and bottom-up sucrose gradients were used to purify schistosomula EVs. Shown are western blot detections of TSP-2 in fractions collected from 13.5 hours centrifuged gradients. Gradients were loaded with material from 11 or 10 ml culture medium from the adult worms or schistosomula, respectively. Western blots shown are representative for 2-3 independent experiments.

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