title: Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations.
abstract: The bioartificial kidney (BAK) aims at improving dialysis by developing 'living membranes' for cells-aided removal of uremic metabolites. Here, unique human conditionally immortalized proximal tubule epithelial cell (ciPTEC) monolayers were cultured on biofunctionalized MicroPES (polyethersulfone) hollow fiber membranes (HFM) and functionally tested using microfluidics. Tight monolayer formation was demonstrated by abundant zonula occludens-1 (ZO-1) protein expression along the tight junctions of matured ciPTEC on HFM. A clear barrier function of the monolayer was confirmed by limited diffusion of FITC-inulin. The activity of the organic cation transporter 2 (OCT2) in ciPTEC was evaluated in real-time using a perfusion system by confocal microscopy using 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP(+)) as a fluorescent substrate. Initial ASP(+) uptake was inhibited by a cationic uremic metabolites mixture and by the histamine H2-receptor antagonist, cimetidine. In conclusion, a 'living membrane' of renal epithelial cells on MicroPES HFM with demonstrated active organic cation transport was successfully established as a first step in BAK engineering.
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.|Department of Physiology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.|Department of Pediatrics, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Jansen
authors.affiliation: Department of Biomaterials Science and Technology, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, The Netherlands.
authors.forename: I E
authors.initials: IE
authors.lastname: De Napoli
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.|Department of Physiology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.|Department of Pediatrics, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Fedecostante
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.|Department of Physiology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.|Department of Pediatrics, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: C M S
authors.initials: CM
authors.lastname: Schophuizen
authors.affiliation: Department of Biomaterials Science and Technology, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, The Netherlands.
authors.forename: N V
authors.initials: NV
authors.lastname: Chevtchik
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.
authors.forename: M J
authors.initials: MJ
authors.lastname: Wilmer
authors.affiliation: Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.
authors.forename: A H
authors.initials: AH
authors.lastname: van Asbeck
authors.affiliation: Department of Cell Biology, Radboud university medical center, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.
authors.forename: H J
authors.initials: HJ
authors.lastname: Croes
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.
authors.forename: J C
authors.initials: JC
authors.lastname: Pertijs
authors.affiliation: Department of Nephrology, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: J F M
authors.initials: JF
authors.lastname: Wetzels
authors.affiliation: Department of Nephrology, Radboud university medical center, Nijmegen, The Netherlands.
authors.forename: L B
authors.initials: LB
authors.lastname: Hilbrands
authors.affiliation: Department of Pediatrics, Radboud university medical center, Nijmegen, The Netherlands.|Department of Pediatric Nephrology &Growth and Regeneration, Catholic University Leuven, Leuven, Belgium.
authors.forename: L P
authors.initials: LP
authors.lastname: van den Heuvel
authors.affiliation: Department of Physiology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.
authors.forename: J G
authors.initials: JG
authors.lastname: Hoenderop
authors.affiliation: Department of Biomaterials Science and Technology, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, The Netherlands.
authors.forename: D
authors.initials: D
authors.lastname: Stamatialis
authors.affiliation: Department of Pharmacology and Toxicology, Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.|Div. Pharmacology, Department of Pharmaceutical Sciences, Utrecht University, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: Masereeuw
mesh_terms: D002412:Cations / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002927:Cimetidine / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D006635:Histamine H2 Antagonists / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D017136:Ion Transport / Q000187:drug effects; D007687:Kidney Tubules, Proximal / Q000166:cytology; D008567:Membranes, Artificial; D008744:Methylamines / Q000737:chemistry / Q000378:metabolism; D027701:Organic Cation Transport Proteins / Q000378:metabolism*; D000075102:Organic Cation Transporter 2; D010539:Permeability / Q000187:drug effects; D011726:Pyridinium Compounds / Q000737:chemistry / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: C517027:4-(4-(dimethylamino)styryl)-1-methylpyridinium iodide; D002412:Cations; D006635:Histamine H2 Antagonists; D008567:Membranes, Artificial; D008744:Methylamines; D027701:Organic Cation Transport Proteins; D000075102:Organic Cation Transporter 2; D011726:Pyridinium Compounds; C491013:SLC22A2 protein, human; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D002927:Cimetidine