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. 2020 May 19;10(5):784. doi: 10.3390/biom10050784

Figure 1.

Figure 1

Co-culture model description. IEC were grown in 12-well transwell inserts until confluency and basolaterally exposed to either non-activated or αCD3/CD28-activated PBMC. Apically, IEC were conditioned with 2′-FL or GF in the presence or absence of CpG, a TLR9 agonist mimicking a bacterial trigger (A). After 24 h incubation, basolateral supernatant was collected to analyze the T-cell mediator release. The IEC were set apart and washed with PBS. Then, fresh medium was added and IEC were kept in incubation for an additional 24 h to study the IEC-derived mediator release (B). Alternatively, IEC were washed with PBS and co-cultured with immature moDC for 48 h (C). Then, the basolateral supernatant was collected where the mediator release was studied. Additionally, the phenotype of moDC after IEC/moDC co-culture was analyzed. Subsequently, conditioned moDC (ccDC) were exposed to naïve T-cells in an allogeneic DC/T-cell assay (D). After 5–6 days incubation, the cytokine release was measured in the supernatant.