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Table 2 Methods for the depletion of EVs in serum additives for cell culture medium

From: Heterogeneity of mesenchymal stem cell-derived extracellular vesicles is highly impacted by the tissue/cell source and culture conditions

Method   References
Ultracentrifugation 120,000 g, 18 h, 4 °C, SW32 Ti rotor (Beckman Coulter, Brea, CA, USA) [66]
Ultrafiltration Amicon ultra-15 centrifugal filters (UFC910024, 100 K filters and benchtop Merk Millipore Ltd., Tullagreen, Carrigtwohill, Co. Cork, Ireland), 3,000 g, 55 min, 4 °C [62]
Tangential flow filtration (TFF) hollow fiber-modified polyethersulfone (mPES) membrane filter column (area 1,600 cm2, 500 kDa molecular weight cut off) operated on a KR2i TFF System (Repligen, USA) [67]
Commercially available exosome-depleted serum or medium MesenCult™-ACF Plus (STEMCELL Technologies, China); [68]
Exo-FBS™ (System Biosciences, Mountain View, CA, USA); [69]
OxiumTMEXO (patent No. PCT/CL2019/100175); [70]
RoosterCollect EV Pro™ (RoosterBio Inc., Frederick, MD, USA) [71]
Fibrinogen and fibrin depletion Hydrogel formation was facilitated for 4 h at room temperature (RT) followed by overnight incubation at 4 °C. The resulting coagulated medium was heated to 37 °C for 1 h to enable a complete fibrin clotting. Afterward, a collapse was induced by vigorous shaking followed by centrifugation at 3000 g for 10 min at RT. Finally, the clear medium supernatant was filtered through a 0.22 µm filter (Merck Millipore, Billerica, MA, USA) [72]