Nephrotoxicity and Kidney Transport Assessment on 3D Perfused Proximal Tubules.
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.
['Vormann|Marianne K|MK|', 'Gijzen|Linda|L|', 'Hutter|Simon|S|', 'Boot|Lisette|L|', 'Nicolas|Arnaud|A|', 'van den Heuvel|Angelique|A|', 'Vriend|Jelle|J|', 'Ng|Chee Ping|CP|', 'Nieskens|Tom T G|TTG|', 'van Duinen|Vincent|V|', 'de Wagenaar|Bjorn|B|', 'Masereeuw|Rosalinde|R|', 'Suter-Dick|Laura|L|', 'Trietsch|Sebastiaan J|SJ|', 'Wilmer|Martijn|M|', 'Joore|Jos|J|', 'Vulto|Paul|P|', 'Lanz|Henriette L|HL|']
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 D001692:Biological Transport D018929:Cell Culture Techniques D002460:Cell Line D016764:Cell Polarity D002945:Cisplatin D016572:Cyclosporine D004305:Dose-Response Relationship, Drug D004847:Epithelial Cells D006801:Humans D007674:Kidney Diseases D007687:Kidney Tubules, Proximal D056656:Lab-On-A-Chip Devices D049990:Membrane Transport Modulators D026901:Membrane Transport Proteins D046210:Microfluidic Analytical Techniques D010477:Perfusion D010695:Phlorhizin D051297:Sodium-Glucose Transporter 2 D000077203:Sodium-Glucose Transporter 2 Inhibitors D019108:Tight Junctions
nephrotoxicity proximal tubule transepithelial transport apparent permeability (P app) kidney-on-a-chip