Figure 3

Precisely-controlled cytokine-induced colon barrier disruption model generated using high-throughput TEER screening. (a) Colon micro-tissues were grown in devices for 4 days in PM followed by 3 days in DM before being treated with cytokine doses for 24 h. Over the 24 h treatment window, TEER was regularly measured at 2, 4, 6, 8, 12, and 24 h post treatment. After 24 h, cytokine doses were washed out and replaced with cytokine-free differentiation media. Micro-tissue barrier function was tracked for 2 additional days via TEER measurements. (b) TEER values measured during the damage time course at 2 h intervals, and every 24 h during recovery. (c) Barrier disruption in response to combination dose curve of TNF-α (0.14–100 ng/mL) and IFN-γ (1.6–1000 U/mL). Heat map colors indicate relative TEER and white font reports exact TEER for each well. One representative experiment is reported during cytokine treatment (12 h) and after recovery (72 h) for two separate colon donors (Donor A, Donor B). (d) Dose curves were generated for TNF-α (left) and IFN-γ (right) for each of the sensitizing concentrations of IFN-γ and TNF-α. Data are displayed from Donor A after recovery (72 h after cytokine addition; 48 h after cytokine removal), where each data point was normalized to the pre-treatment TEER for comparison across wells.