Vascular cell basal media

EA.hy926, HEK293, HepG2, MCF7, and MDA MB231 cells, all purchased from the American Type Culture Collection (Manassas, VA, USA), were grown in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 10% heat-inactivated FBS and 1× antibiotic-antimycotic solution (A5955; Sigma-Aldrich Canada, Oakville, ON, Canada). EA.hy926 cells were cultured for 7–20 days with media changed every second day to achieve 100% confluent cells [32 (link)]. Non-confluent (10–20%) cells were obtained 24 h after 1:4 subculture of a confluent plate of cells. THP-1 cells were grown in RPMI-1640 medium supplemented with 0.05 mM 2-mercaptoethanol and 10% FBS [33 (link)]. Pooled human umbilical artery endothelial cells (HUAECs) were purchased commercially (Catalog number: C-12202; Cedarlane, Burlington, ON, Canada) and grown in Vascular Cell Basal Media (Catalog number: PCS-100-030; ATCC) supplemented with Endothelial Cell Growth Kit-VEGF (Catalog number: PCS-100-041; ATCC). The HUAECs were subcultured at 80% confluency and used within 3–5 passages.

Human HCC cell lines SNU-398 and SK-HEP-1, and Human umbilical vein endothelial cell (HUVEC) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Human HCC cell line Huh7 was purchased from National Collection of Authenticated Cell Cultures of Chinese Academy of Sciences (Shanghai, China). SNU-398 was maintained in RPMI 1640 medium (Invitrogen, Carlsbad, CA USA) added with 10% fetal bovine serum (FBS, Invitrogen). SK-HEP-1 was maintained in Eagle’s Minimum Essential Medium (Invitrogen) added with 10% FBS. HUVEC was maintained in Vascular Cell Basal Media (ATCC) supplemented with Endothelial Cell Growth Kit (ATCC). Huh7 was maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) added with 10% FBS. All cells were cultured at 37 °C containing 5% CO₂ under normoxia. Under hypoxia, cells were cultured at 37 °C containing 1% O₂, 94% N₂, and 5% CO₂. The cells were authenticated using STR profiles. All cells were routinely tested as mycoplasma-free.