A morphological and functional comparison of proximal tubule cell lines established from human urine and kidney tissue
Jansen, J.; Schophuizen, C M S; Wilmer, M J; Lahham, S H M; Mutsaers, H A M; Wetzels, J F M; Bank, R A; van den Heuvel, L P; Hoenderop, J G; Masereeuw, R
(2014) Experimental Cell Research, volume 323, issue 1, pp. 87 - 99
(Article)
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
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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.
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Keywords: ATP-Binding Cassette Transporters, Bioartificial Organs, Cadherins, Cell Adhesion Molecules, Cell Culture Techniques, Cell Line, Collagen Type I, Fibronectins, Humans, Inulin, Kidney Tubules, Proximal, Kidneys, Artificial, Multidrug Resistance-Associated Proteins, Neoplasm Proteins, Octamer Transcription Factor-2, P-Glycoprotein, Tissue Engineering, Transendothelial and Transepithelial Migration, Urine
ISSN: 0014-4827
Publisher: Academic Press Inc.
Note: Copyright © 2014 Elsevier Inc. All rights reserved.
(Peer reviewed)