Non malignant human hepatocytes hh

HepG2 and PLC-PRF5 cells were obtained from American Type Culture Collection (Manassas, VA), and cultured in DMEM high glucose medium (HyClone Laboratories, Logan, UT), containing 10% fetal bovine serum and 1% antibiotic–antimycotic (Invitrogen, Grand Island, NY), at 37 °C with 5% CO2. Non-malignant human hepatocytes HH were obtained from Sciencell and cultured as recommended by the supplier. For all studies with extracellular vesicles, vesicle depleted medium was prepared by centrifuging cell-culture medium at 100,000g overnight to spin down any pre-existing vesicle content. Camptothecin and doxorubicin were obtained from Sigma-Aldrich (St. Louis, MO), and sorafenib was obtained from Selleck (Houston, TX). Compounds were dissolved in 100% DMSO (Sigma–Aldrich, St. Louis, MO) and diluted with culture media to the desired concentration with a final DMSO concentration of 0.1%. DMSO 0.1% (v/v) was used as a solvent control.