A morphological and functional comparison of proximal tubule cell lines established from human urine and kidney tissue

Exp Cell Res. 2014 Apr 15;323(1):87-99. doi: 10.1016/j.yexcr.2014.02.011. Epub 2014 Feb 18.

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.

Keywords: Bioartificial kidney; ECM; Regenerative nephrology; Transport.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / antagonists & inhibitors
  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / biosynthesis
  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism
  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters / biosynthesis
  • ATP-Binding Cassette Transporters / metabolism
  • Bioartificial Organs*
  • Cadherins / biosynthesis
  • Cell Adhesion Molecules / biosynthesis
  • Cell Culture Techniques
  • Cell Line
  • Collagen Type I / biosynthesis
  • Collagen Type I / metabolism
  • Fibronectins / biosynthesis
  • Humans
  • Inulin / metabolism
  • Kidney Tubules, Proximal / cytology*
  • Kidneys, Artificial*
  • Multidrug Resistance-Associated Proteins / biosynthesis
  • Multidrug Resistance-Associated Proteins / metabolism
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / metabolism
  • Octamer Transcription Factor-2 / antagonists & inhibitors
  • Octamer Transcription Factor-2 / biosynthesis
  • Octamer Transcription Factor-2 / metabolism
  • Tissue Engineering
  • Transendothelial and Transepithelial Migration / physiology
  • Urine / cytology*

Substances

  • ABCC4 protein, human
  • ABCG2 protein, human
  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters
  • Cadherins
  • Cell Adhesion Molecules
  • Collagen Type I
  • FN1 protein, human
  • Fibronectins
  • Multidrug Resistance-Associated Proteins
  • Neoplasm Proteins
  • Octamer Transcription Factor-2
  • kalinin
  • Inulin