2019-11-13, Prestigiacomo, Vincenzo, Weston, Anna, Suter-Dick, Laura

Chronic liver damage can lead to fibrosis, encompassing hepatocellular injury, activation of Kupffer cells (KC), and activation of hepatic stellate cells (HSC). Inflammation and TGF-β1 are known mediators in the liver fibrosis adverse outcome pathway (AOP). The aim of this project was to develop a suitable rodent cell culture model for the investigation of key events involved in the development of liver fibrosis, specifically the responses to pathophysiological stimuli such as TGF-β1 and LPS-triggered inflammation. We optimized a single step protocol to purify rat primary hepatocytes (Hep), HSC and KC cells to generate 3D co-cultures based on the hanging drop method. This primary multicellular model responded to the profibrotic cytokine TGF-β1 (1 ng/mL) with signs of hepatocellular damage, inflammation and ultimately HSC activation (increase in αSMA expression). LPS elicited an inflammatory response characterized by increased expression of cytokines. 3D-monocultures comprising only Hep displayed different responses, underlying that parenchymal and non-parenchymal cells need to be present in the system to recapitulate fibrosis. The data also suggest that pre-activated HSC may reverse to a quiescent phenotype in 3D, probably due to the more physiological conditions.

2018-11, Suter-Dick, Laura

2018-10, Messner, Catherine, Prestigiacomo, Vincenzo, Mannino, Salvatore, Mauch, Linda, Suter-Dick, Laura

2018-08, Vormann, Marianne K., Gijzen, Linda, Hutter, Simon, Boot, Lisette, Nicolas, Arnaud, van den Heuvel, Angelique, Vriend, Jelle, Ng, Chee Ping, Nieskens, Tom T.G., van Duinen, Vincent, de Wagenaar, Bjorn, Masereeuw, Rosalinde, Suter-Dick, Laura, Trietsch, Sebastian J., Wilmer, Martijn, Joore, Jos, Vulto, Paul, Lanz, Henriette

Proximal tubules in the kidney play a crucial role in reabsorbing and eliminating substrates from the body into the urine, leading to high local concentrations of xenobiotics. This makes the proximal tubule a major target for drug toxicity that needs to be evaluated during the drug development process. Here, we describe an advanced in vitro model consisting of fully polarized renal proximal tubular epithelial cells cultured in a microfluidic system. Up to 40 leak-tight tubules were cultured on this platform that provides access to the basolateral as well as the apical side of the epithelial cells. Exposure to the nephrotoxicant cisplatin caused a dose-dependent disruption of the epithelial barrier, a decrease in viability, an increase in effluent LDH activity, and changes in expression of tight-junction marker zona-occludence 1, actin, and DNA-damage marker H2A.X, as detected by immunostaining. Activity and inhibition of the efflux pumps P-glycoprotein (P-gp) and multidrug resistance protein (MRP) were demonstrated using fluorescence-based transporter assays. In addition, the transepithelial transport function from the basolateral to the apical side of the proximal tubule was studied. The apparent permeability of the fluorescent P-gp substrate rhodamine 123 was decreased by 35% by co-incubation with cyclosporin A. Furthermore, the activity of the glucose transporter SGLT2 was demonstrated using the fluorescent glucose analog 6-NBDG which was sensitive to inhibition by phlorizin. Our results demonstrate that we developed a functional 3D perfused proximal tubule model with advanced renal epithelial characteristics that can be used for drug screening studies.

2019, Suter-Dick, Laura, Greim, Helmut, Snyder, Robert

2018-11, Yan, Lu, Zenker, Armin, Suter-Dick, Laura

2018-10, Suter-Dick, Laura

2018-11, Messner, Catherine, Mauch, Linda, Mannino, Salvatore, Prestigiacomo, Vincenzo, Suter-Dick, Laura

2018-11, Suter-Dick, Laura

2018-09, Gaiser, Carine, Weston, Anna, Suter-Dick, Laura