Figure 3
From: Bioengineered kidney tubules efficiently excrete uremic toxins
Bioengineered renal tubules show transepithelial barrier function and polarized characteristics.
FITC-inulin leakage was measured in matured ciPTEC-OAT1 seeded on coated HFM. (a) Schematic presentation of the experimental set-up in the absence (upper panel) or presence (lower panel) of cells. (b) Representative eal-time images of the two different conditions are shown; unseeded (square) and seeded (circle) HFM. In the absence of cells, a bright green fluorescent signal was detected at the apical membrane, whereas in the presence of cells the signal was clearly attenuated. Representative real-time images of the two different conditions were shown. (c) Semi-quantification of FITC-inulin diffusion in the absence (square) and the presence of ciPTEC-OAT1 (circle) on HFM. Seeded HFM showed significantly less FITC-inulin leakage demonstrating monolayer tightness. From each single replicate 4 different regions in focus were analyzed. (d) The expression of ZO-1 (green) was demonstrated in a homogenous cell monolayer cultured on HFM. Scale bar: 10 μm. (e–g) The OAT1, BCRP and MRP4 mRNA expression levels were investigated in matured ciPTEC-OAT1 cultured in well plates and on HFM. Gene expression levels were normalized to expression levels of the reference gene GADPH (h) and were expressed as fold increase when cultured on HFM (black) compared to matured cells in well plates (white). Data are shown as mean ± S.E.M. of three independent experiments performed, at least, in duplicate. ***p < 0.001, using an unpaired t test.
