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Properties of a polarized primary culture from rat renal inner medullary collecting duct (IMCD) cells
Authors:Birgit Ruhfus  H. Günther Bauernschmitt  Rolf K. H. Kinne
Affiliation:(1) Abteilung Epithelphysiologie, Max-Planck-Institut für molekulare Physiologie, Rheinlanddamm 201, 44139 Dortmund, Germany;(2) Abteilung Epithelphysiologie, Max-Planck-Institut für molekulare Physiologie, Postfach 10 26 64, 44026 Dortmund, Germany
Abstract:
Summary A primary culture from rat renal IMCD cells was established to investigate the permeability characteristics of the luminal and contraluminal plasma membranes of the papillary collecting duct in vitro. Freshly isolated IMCD cells were grown on filters in a special “epithelial cell” medium. Confluency was proved with an epithelial volt/ohm meter. After 7 d of culture the transepithelial resistance reached more than 1000 Ω×cm2. A polarization of the cells with regard to a basolateral localization of a lactate efflux system, and an l-alanine transport system was achieved. The hypotonicity-activated release systems for the organic osmolytes sorbitol and betaine were also located basolaterally, whereas taurine, glycerophosphorylcholine, and myo-inositol left the cells at both cell poles but with different capacity. Morphological observations revealed also that the monolayer was well differentiated. Thus, a model of a renal collecting duct epithelium was established which can be used to analyze polarized and differentiated transport processes across the epithelial cells and their plasma membranes.
Keywords:collecting duct  amino acid transport  organic osmolytes  cell polarity
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