title: A morphological and functional comparison of proximal tubule cell lines established from human urine and kidney tissue.
abstract: Promising renal replacement therapies include the development of a bioartificial kidney using functional human kidney cell models. In this study, human conditionally immortalized proximal tubular epithelial cell (ciPTEC) lines originating from kidney tissue (ciPTEC-T1 and ciPTEC-T2) were compared to ciPTEC previously isolated from urine (ciPTEC-U). Subclones of all ciPTEC isolates formed tight cell layers on Transwell inserts as determined by transepithelial resistance, inulin diffusion, E-cadherin expression and immunocytochemisty. Extracellular matrix genes collagen I and -IV α1 were highly present in both kidney tissue derived matured cell lines (p<0.001) compared to matured ciPTEC-U, whereas matured ciPTEC-U showed a more pronounced fibronectin I and laminin 5 gene expression (p<0.01 and p<0.05, respectively). Expression of the influx carrier Organic Cation Transporter 2 (OCT-2), and the efflux pumps P-glycoprotein (P-gp), Multidrug Resistance Protein 4 (MRP4) and Breast Cancer Resistance Protein (BCRP) were confirmed in the three cell lines using real-time PCR and Western blotting. The activities of OCT-2 and P-gp were sensitive to specific inhibition in all models (p<0.001). The highest activity of MRP4 and BCRP was demonstrated in ciPTEC-U (p<0.05). Finally, active albumin reabsorption was highest in ciPTEC-T2 (p<0.001), while Na(+)-dependent phosphate reabsorption was most abundant in ciPTEC-U (p<0.01). In conclusion, ciPTEC established from human urine or kidney tissue display comparable functional PTEC specific transporters and physiological characteristics, providing ideal human tools for bioartificial kidney development.
authors.affiliation: Department of Pharmacology and Toxicology, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Physiology, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Pediatrics, Radboud University Medical Center, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Jansen
authors.affiliation: Department of Pharmacology and Toxicology, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Physiology, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Pediatrics, Radboud University Medical Center, The Netherlands.
authors.forename: C M S
authors.initials: CMS
authors.lastname: Schophuizen
authors.affiliation: Department of Pharmacology and Toxicology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: M J
authors.initials: MJ
authors.lastname: Wilmer
authors.affiliation: Department of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
authors.forename: S H M
authors.initials: SHM
authors.lastname: Lahham
authors.affiliation: Department of Pharmacology and Toxicology, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Physiology, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Pediatrics, Radboud University Medical Center, The Netherlands.
authors.forename: H A M
authors.initials: HAM
authors.lastname: Mutsaers
authors.affiliation: Department of Nephrology, Radboud University Medical Center, The Netherlands.
authors.forename: J F M
authors.initials: JFM
authors.lastname: Wetzels
authors.affiliation: Department of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
authors.forename: R A
authors.initials: RA
authors.lastname: Bank
authors.affiliation: Department of Pediatrics, Radboud University Medical Center, The Netherlands; Department of Pediatrics, Catholic University Leuven, Leuven, Belgium.
authors.forename: L P
authors.initials: LP
authors.lastname: van den Heuvel
authors.affiliation: Department of Physiology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: J G
authors.initials: JG
authors.lastname: Hoenderop
authors.affiliation: Department of Pharmacology and Toxicology, Radboud University Medical Center, Nijmegen, The Netherlands. Electronic address: roos.masereeuw@radboudumc.nl.
authors.forename: R
authors.initials: R
authors.lastname: Masereeuw
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis / Q000378:metabolism; D021522:Bioartificial Organs; D015820:Cadherins / Q000096:biosynthesis; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D018929:Cell Culture Techniques; D002460:Cell Line; D024042:Collagen Type I / Q000096:biosynthesis / Q000378:metabolism; D005353:Fibronectins / Q000096:biosynthesis; D006801:Humans; D007444:Inulin / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology*; D007688:Kidneys, Artificial; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis / Q000378:metabolism; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000378:metabolism; D050813:Octamer Transcription Factor-2 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism; D023822:Tissue Engineering; D057705:Transendothelial and Transepithelial Migration / Q000502:physiology; D014556:Urine / Q000166:cytology*; D000097794:Kalinin
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D015820:Cadherins; D015815:Cell Adhesion Molecules; D024042:Collagen Type I; D005353:Fibronectins; D007444:Inulin; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009363:Neoplasm Proteins; D050813:Octamer Transcription Factor-2; D000097794:Kalinin; C073492:ABCC4 protein, human; C116614:ABCG2 protein, human; C067625:FN1 protein, human
keywords: Bioartificial kidney; ECM; Regenerative nephrology; Transport