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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/8554

Title: A primary culture of mouse proximal tubular cells, established on collagen-coated membranes
Authors: TERRYN, Sara
Jouret, Francois
VANDENABEELE, Frank
SMOLDERS, Inge
MOREELS, Marjan
Devuyst, Olivier
STEELS, Paul
VAN KERKHOVE, Emmy
Issue Date: 2007
Publisher: AMER PHYSIOLOGICAL SOC
Citation: AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, 293(2). p. F476-F485
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 gamma- 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, B-3590 Diepenbeek, Belgium. Univ Hasselt, Histol Lab, B-3590 Diepenbeek, Belgium. Transnatl Univ Limburg, Diepenbeek, Belgium. Inst Biomed, Diepenbeek, Belgium. Ctr Environm Sci, Diepenbeek, Belgium. Catholic Univ Louvain, Div Nephrol, Brussels, Belgium.
URI: http://hdl.handle.net/1942/8554
DOI: 10.1152/ajprenal.00363.2006
ISI #: 000248459000005
ISSN: 0363-6127
Category: A1
Type: Journal Contribution
Validation: ecoom, 2008
Appears in Collections: Research publications

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