Hepa 1 6 cells crl 1830

HepG2 cells (HB-8065, American Type Culture Collection, Manassas, VA, USA) were cultured in DMEM supplemented with 10% FBS, 1% L-glutamine (200 mM), penicillin (100 U/mL), and streptomycin (100 µg/mL), at 37 °C, 5% (v/v) CO₂. HUVECs cells (C-12200, PromoCell GmbH, Heidelberg, Germany) were cultured in EGM2 supplemented with Supplement Mix/Endothelial (PromoCell GmbH, Heidelberg, Germany), penicillin (100 U/mL) and streptomycin (100 µg/mL), at 37 °C, 5% (v/v) CO₂. HUVECs were cultured in flasks pre coated with 2% collagen solution (4 mg/mL) to a maximum of 8 passages as specified by the supplier (15 doublings). Hepa 1–6 cells (CRL-1830, American Type Culture Collection, Manassas, Virginia, USA) were cultured in DMEM supplemented with 10% FBS, 1% L-glutamine (200 mM), penicillin (100 U/mL), and streptomycin (100 µg/mL), at 37 °C, 5% (v/v) CO₂.

HSCs were isolated from WT and Cygb^−/− mice using the pronase-collagenase digestion method as described previously66 (link) and were cultured on uncoated-plastic dishes (BD Falcon, Franklin Lake, NY, USA) or glass chamber slides (Thermo Fisher Scientific, Waltham, MA, USA) in DMEM (Sigma-Aldrich) supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA) and antibiotics (100 U/ml penicillin and 100 μg/ml streptomycin) at 37 °C in a 5% CO₂/95% room air. Mouse hepatoma Hepa 1–6 cells (CRL-1830) obtained from American Type Culture Collection (Manassas, VA, USA) were maintained on uncoated-plastic culture plates (BD Falcon) in DMEM supplemented with 10% FBS and antibiotic.