4.2. IEC Model

IEC were seeded (50.000 cells/well) in 48-well plates (Costar Corning Incorporated, New York, NY, USA) and grown until confluency in Mc Coy 5A medium. Medium was refreshed every 2–3 days. When IEC reached confluency, cells were pre-incubated with 0.5% (w/v; 5 mg/mL) 2′FL solutions (>90% pure, produced by microbial fermentation) for 24 h. After the pre-incubation period, medium was removed and IEC were stimulated with 10 µg/mL high-molecular weight pIC (Invivogen, San Diego, CA, USA) either naked or complexed with Lyovec™ (LV) (Invivogen), alone or in the presence of 2′FL, for 20 h after which, the supernatant was collected and stored for cytokine analysis.