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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/2015

Title: A primary culture of mouse proximal tubule cells, established on collagen-coated membranes
Authors: TERRYN, Sara
Jouret, F
VANDENABEELE, Frank
SMOLDERS, Inge
MOREELS, Marjan
Devuyst, O
STEELS, Paul
VAN KERKHOVE, Emmy
Issue Date: 2006
Publisher: OXFORD UNIV PRESS
Citation: NEPHROLOGY DIALYSIS TRANSPLANTATION, 21. p. 59-59
Abstract: A simple method is described to establish primary cultures of kidney proximal tubule cells (PTC) on membranes. The permeable membranes represent a unique culture surface, allowing a high degree of differentiation since both apical and basolateral membranes are accessible for medium. Proximal tubule (PT) segments from collagenase-digested mouse renal cortices were grown for 7 days, by which time cells were organized as a confluent monolayer. Electron microscopic evaluation revealed structurally polarized epithelial cells with numerous microvilli, basolateral invaginations, and apical tight junctions. Immunoblotting for markers of distinct parts of the nephron demonstrated that these primary cultures only expressed PT-specific proteins. Moreover immunodetection of distinct components of the receptor-mediated endocytic pathway and uptake of FITC-albumin indicated that these cells expressed a functional endocytotic apparatus. In addition, primary cultures possessed the PT-brush border enzymes, alkaline phosphatase, and γ-glutamyl-transferase, and a phloridzin-sensitive sodium-dependent glucose transport at their apical side. Electrophysiological measurements show that the primary cultured cells have a low transepithelial resistance and high short-circuit current that was completely carried by Na+ similar to a leaky epithelium like proximal tubule cells. This novel method established well-differentiated PTC-cultures.
Notes: Univ Hasselt, Lab Cell Physiol, Diepenbeek, Belgium. Univ Hasselt, Histol Lab, Diepenbeek, Belgium. Univ Catholique Louvain, Div Nephrol, B-1200 Brussels, Belgium.
URI: http://hdl.handle.net/1942/2015
DOI: 10.1152/ajprenal.00363.2006
ISI #: 000239919000161
ISSN: 0931-0509
Category: M
Type: Journal Contribution
Appears in Collections: Research publications

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