EV-depleted medium was prepared as previously described [12 (link),13 (link),96 (link)]. In summary, FBS was filtered using Amicon® Ultra-15 centrifugal filters with a 100 kDa cut-off (Merk KGaA, Darmstadt, Germany) at 3000× g for 55 min. The EV-depleted FBS was 90% depleted of nanoparticles and used at a 10% concentration to supplement all complete culture media specific to each cell type mentioned above.
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