To assess the effect of ethanol (EtOH; 10 or 50 mM), lipopolysaccharide (LPS; 50 ng/ml) or ATP (5 mM), these compounds were added to DMEM in the absence of serum, but in the presence of 1 mg/ml bovine serum albumin. After 24 h of LPS, ATP or ethanol treatments, cells were harvested by trypsinization, centrifuged and used for specific determinations. The medium was used for cytokine determination.
