2.6. In vitro cortisol determination assay

H295R cells were seeded in 24 well cell culture plates at a density of 75,000 cells/ml and incubated for 24h before treatments. Following an incubation with water (control), TBP at 500 μg/ml or caffeine at 10 mM in serum-free medium for 24 h, measurements of cortisol concentrations from the cell culture medium were performed using the DetectX® Cortisol assay kit (Arbor Assays, Ann Arbor, MI, USA) according to the manufacturer's instructions.