title: Upscaling of a living membrane for bioartificial kidney device.
abstract: The limited removal of metabolic waste products in dialyzed kidney patients leads to high morbidity and mortality. One powerful solution for a more complete removal of those metabolites might be offered by a bioartificial kidney device (BAK), which contains a hybrid "living membrane" with functional proximal tubule epithelial cells (PTEC). These cells are supported by an artificial functionalized hollow fiber membrane (HFM) and are able to actively remove the waste products. In our earlier studies, conditionally immortalized human PTEC (ciPTEC) showed to express functional organic cationic transporter 2 (OCT2) when seeded on small size flat or hollow fiber polyethersulfone (PES) membranes. Here, an upscaled "living membrane" is presented. We developed and assessed the functionality of modules containing three commercially available MicroPES HFM supporting ciPTEC. The HFM were optimally coated with L-Dopa and collagen IV to support a uniform and tight monolayer formation of matured ciPTEC under static culturing conditions. Both abundant expression of zonula occludens-1 (ZO-1) protein and limited diffusion of FITC-inulin confirm a clear barrier function of the monolayer. Furthermore, the uptake of 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP+), a fluorescent OCT2 substrate, was studied in absence and presence of known OCT inhibitors, such as cimetidine and a cationic uremic solutes mixture. The ASP+uptake by the living upscaled membrane was decreased by 60% in the presence of either inhibitor, proving the active function of OCT2. In conclusion, this study presents a successful upscaling of a living membrane with active organic cation transport as a support for BAK device.
authors.affiliation: Department of Biomaterials Science and Technology, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Enschede, The Netherlands.
authors.forename: Natalia Vladimirovna
authors.initials: NV
authors.lastname: Chevtchik
authors.affiliation: Department of Pharmaceutical Sciences, UIPS Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Michele
authors.initials: M
authors.lastname: Fedecostante
authors.affiliation: Department of Pharmaceutical Sciences, UIPS Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Jitske
authors.initials: J
authors.lastname: Jansen
authors.affiliation: Department of Pharmaceutical Sciences, UIPS Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Milos
authors.initials: M
authors.lastname: Mihajlovic
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Radboudumc, Nijmegen, The Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Wilmer
authors.affiliation: eXcorLab GmbH, Industrie Center Obernburg, Obernburg, Germany.
authors.forename: Marieke
authors.initials: M
authors.lastname: Rüth
authors.affiliation: Department of Pharmaceutical Sciences, UIPS Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Rosalinde
authors.initials: R
authors.lastname: Masereeuw
authors.affiliation: Department of Biomaterials Science and Technology, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, Enschede, The Netherlands. Electronic address: d.stamatialis@utwente.nl.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Stamatialis
mesh_terms: D001692:Biological Transport; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D007444:Inulin / Q000031:analogs & derivatives / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology; D007688:Kidneys, Artificial; D008567:Membranes, Artificial; D027701:Organic Cation Transport Proteins / Q000378:metabolism
chemical_list: C584353:FITC-inulin; D008567:Membranes, Artificial; D027701:Organic Cation Transport Proteins; D007444:Inulin; D016650:Fluorescein-5-isothiocyanate
keywords: Bioartificial kidney device; Conditionally immortalized human renal proximal tubule epithelial cells; Functionalized hollow fiber membranes; Living membrane; Monolayer of functional cells