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

J. Jansen, Ilaria De Napoli, M. Fedecostante, C.M.S. Schophuizen, Natalia Chevtchik, M. Wilmer, A.H. van Asbeck, H.J. Croes, J.C. Pertijs, F.J.M. Wetzels, L.B Hilbrands, L.P. van der Heuvel, J.G. Hoenderop, Dimitrios Stamatialis, R. Masereeuw

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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.
Original languageEnglish
Article number16702
Number of pages12
JournalScientific reports
Volume5
DOIs
Publication statusPublished - 16 Nov 2015

Keywords

  • METIS-313824
  • IR-99626

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