Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations

• DOI: 10.1038/srep16702
• Publication Date: 2015-11-16T10:01:40Z
• Authors: J. Jansen; I. E De Napoli; M. Fedecostante; C. M. S. Schophuizen; N. V. Chevtchik; M. J. Wilmer; A. H. van Asbeck; H. J. Croes; J. C. Pertijs; J. F. M. Wetzels; L. B. Hilbrands; L. P. van den Heuvel; J. G. Hoenderop; D. Stamatialis; R. Masereeuw
• Keywords: 0301 basic medicine; Organic Cation Transport Proteins; Pyridinium Compounds; Article; Permeability; Cell Line; Tight Junctions; Kidney Tubules, Proximal; Methylamines; 03 medical and health sciences; Radboudumc 16: Vascular damage RIHS: Radboud Institute for Health Sciences; Cations; Cell Adhesion; Humans; Radboudumc 11: Renal disorders RIHS: Radboud Institute for Health Sciences; 0303 health sciences; Ion Transport; Radboudumc 19: Nanomedicine RIMLS: Radboud Institute for Molecular Life Sciences; Organic Cation Transporter 2; Epithelial Cells; Membranes, Artificial; Radboudumc 11: Renal disorders RIMLS: Radboud Institute for Molecular Life Sciences; Immunohistochemistry; Histamine H2 Antagonists; Zonula Occludens-1 Protein; Cimetidine

AbstractThe 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.