Hepg2 human hepatocarcinoma

The HepG2 human hepatocarcinoma and the human neuroblastoma SH-SY5Y cell lines were obtained from the European Collection of Cell Cultures (Health Protection Agency, London, UK) (ECACC Ref.-85011430 and Ref.-94030304, respectively). The HepG2 cell line is a model that reproduces human hepatocyte and has been widely used in the evaluation of the antioxidant effects of various natural compounds [23 (link)]. On the other hand, SY-SY5Y is commonly used as an in vitro model of neural function and neural differentiation [25 (link)]. HepG2 cells were grown in EMEM (Eaglee’s Minimum Essential Medium) and SH-SY5Y in Ham’s F-12: EMEM (EBSS) 1:1, supplemented both with 2 mM glutamine, 1% non-essential amino acids, inactivated fetal bovine serum (FBS) (HyClone, Logan, UT, USA) (10% for HepG2 and 15% for SH-SY5Y), and 1% antibiotics (10,000 units of penicillin and 10 mg/mL of streptomycin). Both HEPG2 and SY-SY5Y were cultured at 37 °C in a humidified 5% CO₂ atmosphere. For antioxidant assays, cells were incubated in a medium supplemented with only 1% FBS in order to prevent cytotoxic artefacts from forming as a result of the possible interaction between FBS components and phenolic compounds [26 (link)]. The chemicals were obtained from Sigma (Merck KGaA, Darmstadt, Germany).

The HepG2 human hepatocarcinoma and the RAW 264.7 murine macrophage cell lines were obtained from the European Collection of Cell Cultures (Health Protection Agency, London, UK) (ECACC ref. 85011430 and ref. 91062702, respectively).
In both cases, cells were grown in EMEM (Eagle’s Minimum Essential Medium) supplemented with 2 mM glutamine, 1% non-essential amino acids, 10% foetal bovine serum (FBS), and 1% antibiotics (10 mg/mL streptomycin and 10,000 U penicillin). They were kept in an incubator at 37 °C in an atmosphere of 5% CO₂ in air. The state of the culture was regularly checked every 24 h by observation under a microscope.