Hepg2 human hepatoma cells

HepG2 human hepatoma cells (ATCC, Manassas, VA, USA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Hyclone, Logan, UT, USA, SH30021.01) supplemented with 10% FBS (Gibco, 16000044) and 1% antibiotics–antimycotics (Gibco, 15250062) at 37 °C in a humidified atmosphere containing 5% CO₂. For infection of mito-Catalase adenovirus, HepG2 cells were seeded in 12-well plates and incubated with the adenovirus the following day for 24 h. Mouse primary hepatocyte isolation was performed as described previously [4 (link)].

The testicular cancer cell lines, SEM-1 and TCAM-2 (a kind gift from Prof. A.L. Epstein, CA, USA), and HepG2 human hepatoma cells (American Type Culture Collection, Manassas, VA, USA) were cultured in an RPMI-1640 culture medium, supplemented with 10% fetal bovine serum, 1% glutammine, and 1 mg/mL penicillin/streptomycin (Sigma Aldrich, Milano, Italy). All of the cell lines were cultured in 100 mm dishes, and were kept incubated at 37 °C in an atmosphere of 5% CO₂. All of the experiments were performed after 12 h of cell synchronization in serum-free media (SFM). The OLE was obtained from Sigma Aldrich (cat. N. O8889, St. Louis, MO, USA), and was dissolved in double-distilled water and diluted in RPMI-1640 media before use.

HepG2 human hepatoma cells were purchased from the American Type Culture Collection (Manassas, VA, USA) and cultured at 37 °C/5% CO₂ in Minimum Essential Medium (MEM; Cat. LM001-01, WELGENE, Gyeongsan, Republic of Korea) containing 10% fecal bovine serum (FBS; Cat. 10082147, Gibco, Waltham, MA, USA) and 1% penicillin–streptomycin (P/S; Cat. 15140122, Gibco, Waltham, MA, USA). We used 10 passages of HepG2 cells.

HepG2 human hepatoma cells were obtained from the American type Culture Collection (ATCC). Cells were maintained in 10 % Growth Medium (GM), consisting of Dulbecco’s Modified Eagle’s Medium (high glucose with 4500 mg/L glucoseand sodium bicarbonate without L-glutamine; DMEM; Sigma) supplemented with 10 % (v/v) fetal Bovine serum (FBS; Sigma), antibiotics (penicillin 100U/ml, 100 μg/mlstreptomycin sulphate) and 2 mM L-glutamine (Sigma). Adherent HepG2 and McARH-7777 cells were maintained in monolayer culture in 75 cm² flasks in 10 % GM and incubated at 37 °C, in 5 % CO₂ for HepG2 and 10 % CO₂for McARH-7777 cells. Fresh GM was added every 2 days and cells were sub-cultured once a week by trypsinisation when the cells were 70 %–80 % confluent. The medium was removed by aspiration and the cells washed with 3 ml 1x EDTA/saline. One milliliter of 1x trypsin solution was added and the flask incubated at 37 °C for 2–3 min. Ten milliliters of 10 % GM was added and the cells were harvested by centrifugation at 900 × gfor 5 min. The cell pellet was resuspended in fresh 10 % GM. Cells were split 1:2 to 1:7 into 75cm² flasks for growth or into 6-well plates (9.6 cm²/well; IWAKI) for experiments. Hesperetin (purity: >98 %, was purchased from Sigma).