Figure 6
Epithelial cell monolayer formation and transepithelial barrier function following co-culture with PBMC. (a) Transwell® inserts and microPES hydrophilic flat membrane used for the indirect co-culture of ciPTEC-T1 and PBMC. (b) ZO-1 expression (red) in ciPTEC-T1 cultured on double-coated microPES membranes after 7 day maturation at 37 °C, nuclei stained with DAPI (blue) (25X magnification). (c) Transepithelial inulin-FITC diffusion in absence of cells, in presence of double-coating (L-DOPA 2 mg/ml and collagen IV 25 μg/ml), untreated cells and cells exposed to cisplatin 50 μM for 24 h and (d) Iinulin-FITC diffusion across membranes containing ciPTEC-T1 after 48 h of co-culture with PBMC in the presence of several stimulatory conditions, including plasma from ESRD patients. Diffusion is shown as pmol·min−1·cm−2 (mean ± SEM). Two independent experiments in triplicate (c) and three independent experiments in duplicate (d) were performed. *p < 0.05, ***p < 0.001, compared to non-coated membranes; #p < 0.05, ##p < 0.01 and ###p < 0.001, compared to coated membranes; $$$p < 0.001, compared to ciPTEC (untreated control; One-way ANOVA, Dunnett’s multiple comparison test).
