Fig. 3.

Endothelial cell presence and TNF-α modify trophoblast migration, FKBPL and Gal-3 expression in a microfluidic chip. In the co-culture set of chips, HUVECs were embedded within the center matrix channel and ACH-3Ps were added to the side channel. a Representative immunofluorescence (IF) images of ACH-3Ps and HUVECs with high expression for FKBPL and CD31, respectively. Nuclei of cells were visualized using DAPI. b Representative IF images of ACH-3Ps invasion across the device (left to right) in the absence or presence of HUVECs and in normal or inflammatory conditions. Cells were IF stained for cytokeratin 7, a marker of trophoblasts, and DAPI. e The number of migrating trophoblast cells from the left side channel were analyzed using ImageJ. c, d ACH-3Ps monoculture chips were also fixed and IF stained for FKBPL, Galectin-3 and DAPI. Chips were treated with TNF-α (10 ng/mL) for 24 or 72 h, with untreated cells as a control. The fold change of f FKBPL expression in ACH-3Ps without HUVECs and h with HUVECs. The fold change of g Gal-3 expression in ACH-3Ps without HUVECs and i with HUVECs Gal-3. Scalebars represent 100 µm. Data plotted as mean fold change ± SEM, ordinary one-way ANOVA or two-way ANOVA with Tukey post hoc test, n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001