Figure 5

Enhancement of intestinal epithelial integrity. 2D enteroids grown on Matrigel coated transwells. The duration of the maximum TEER of 2D enteroids grown can be extended by A addition of CHIR99021 (10 mM) in the culture media from initial seeding for longer than 24 h (control, 24 h), B increasing Ca²⁺ concentration from control concentration 1.05 mM to 2 mM by addition of calcium chloride (duplicate wells) or by C addition of sodium butyrate (NaB; 3, 5, 7 mM; duplicate wells). D NaB and Ca²⁺ rescue epithelial integrity of a leaky gut model of 2D enteroids grown in low calcium media (LCM with 0.26 mM Ca²⁺). The barrier integrity could be rescued by restoring Ca²⁺ concentration to the control concentration (1.05 mM Ca²⁺) or by adding NaB (7 mM) from day 3 of culture. All additions were made every second day, from day 3 of culture, to the apical and basal compartments of the transwell. Statistical significance compared to control wells are *P < 0.05, **0.05 < P > 0.001, ***P < 0.001. Data is presented as the median of 3–5 independent experiments and 95% CI.