Use of cultured tubular cells isolated from human urine for investigation of renal transporter

Clin Nephrol. 2000 Feb;53(2):90-8.


Aim: To facilitate the understanding of the transporter function of human renal tubular cells, we have developed a simple method using primary cultured proximal tubule (PT) cells isolated from voided urine.

Methods: PT cells grown to confluence on glass coverslips could be identified by parallel arrays of spindle cells and hemicyst formation. Brush-border gamma-glutamyl transpeptidase (gammaGTP) activity was histochemically identified. Apical membrane Na+/H+ exchanger (NHE) activity was measured by monitoring changes in intracellular pH (pHi) after an acid load in a single cell level using the pH-sensitive dye 2'7'-bis-(2-carboxyethyl)-5.6'carboxyfluorescein (BCECF).

Results: Amiloride and 5-(N-ethyl-N-isopropyl) amiloride (EIPA) inhibited the NHE activity with half-maximal inhibition values (IC50) of 15.3 and 4.0 microM, respectively. NHE-3 mRNA was detected by the RT-PCR technique in clonally proliferated PT cells.

Conclusion: These results suggest that cultured PT cells isolated from human urine express amiloride-resistant NHE-3 activity on the apical membranes, which can be compared to functional properties of PT in vivo. Our experimental strategy offers a useful experimental approach to investigating human renal tubular transport function in vitro.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Base Sequence
  • Biological Transport
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Child, Preschool
  • DNA Primers
  • Histocytochemistry
  • Humans
  • Infant
  • Infant, Newborn
  • Kidney Tubules, Proximal / cytology*
  • Kidney Tubules, Proximal / metabolism
  • Male
  • Microscopy, Electron
  • Molecular Sequence Data
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Sodium-Hydrogen Exchangers / metabolism
  • Urine / cytology
  • gamma-Glutamyltransferase / metabolism


  • DNA Primers
  • Sodium-Hydrogen Exchangers
  • gamma-Glutamyltransferase