Behavioral changes in H69 cells in quiescent culture by the pathogenic effect of ESP treatment and the diffusion profile of ESPs in a COL1 hydrogel.

(A) Our model predicts that ESP concentrations varied in the region of the biliary ductal plate (red dotted lines) between 1.5 μg/mL to 3.0 μg/mL and 1.7 μg/mL to 0.5 μg/mL after direct and gradient application respectively. Concentration profiles and heat-map images show the ESP concentration along the cell-seeded channel, COL1 hydrogel scaffold and opposite channel. (B) H69 cells sprouting induced by ESP treatment conditions; non-treated control (NC), direct treatment (Direct) and treatment with a gradient (Gradient). Dotted lines (red) and arrows (red) indicate the biliary ductal plate and H69 cell sprouting, respectively. (C) Quantification of the area of H69 cell sprouting into the COL1 hydrogel (area). There was no significant difference between the direct and gradient ESP applications and the NC. Scale bars = 100 μm (B). Student’s t-test was used to analyze significance. Error bars = ± SEM.