Beckman 50.2ti

Unpurified EV solution was further ultracentrifuged at 4 °C at medium speed (10,000 to 20,000× g) for 70 min to pellet large EVs, then at high speed (100,000 to 164,000× g) for 70 min to pellet small EVs. Rotors used were either Beckman 50.2Ti or SW-32.1 (337901/369651 Beckman Coulter France, Villepinte, France).

EVs were isolated by ultracentrifugation from HeLa conditioned culture media (media from 4 dishes/sample), human plasma (250 µL/sample) and human cardiac biopsies (200 mg/sample were minced and incubated in 6 mL PBS 1X containing 1 mg/mL type II collagenase (LS004174, Worthington) at 37 °C for 30 min with agitation). Conditioned culture media, plasma and cardiac minced tissue-containing PBS were centrifuged at 3000 g to remove debris and dead cells. The supernatants were ultracentrifuged at 20,000g (70 min, 4 °C) to pellet large EVs (lEVs), then at 100,000g (70 min, 4 °C) to pellet small EVs (sEVs). Rotors used were either Beckman 50.2 Ti or SW-32.1 (337,901/369651 Beckman Coulter France, Villepinte, France).