Preparation of basolateral membranes that transport p-aminohippurate from primary cultures of rabbit kidney proximal tubule cells

A high degree of functional polarity has been obtained in primary cultures of rabbit kidney proximal tubule cells grown on collagen IV-coated porous membranes. Tight confluency was attained 6 days after seeding and maintained for at least 6 more days, as shown by analysis of paracellular inulin diffusion. From day 6 onward, L-lactate, ammonia, and D-glucose concentration gradient and a pH difference of approximately 1 unit developed between the two nutrient medium compartments. Confluent monolayers expressed organic ion transport properties higher than those formerly reported for other cell models. Transcellular transport of 20 microM tetraethylammonium was directed from basal to apical compartment and was specifically inhibited by mepiperphenidol (1 mM). Unidirectional transport of 2.4 microM p-aminohippurate also occurred from basal to apical compartment, was saturable, and specifically inhibited by probenecid (1 mM). These results suggest that rabbit kidney proximal tubule cells, cultured under the experimental conditions described here, may be a useful model for the in vitro study of highly polarized renal transport processes.

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Changes in expression and subcellular localization of annexin IV in rabbit kidney proximal tubule cells during primary culture

Journal of Cellular Physiology ◽

10.1002/jcp.1041650212 ◽

1995 ◽

Vol 165 (2) ◽

pp. 313-322 ◽

Cited By ~ 5

Author(s):

D. Massey-Harroche ◽

V. Traverso ◽

N. Mayran ◽

V. Francou ◽

A. Vandewalle ◽

...

Keyword(s):

Subcellular Localization ◽

Proximal Tubule ◽

Primary Culture ◽

Rabbit Kidney ◽

Proximal Tubule Cells ◽

Kidney Proximal Tubule ◽

Tubule Cells ◽

Annexin Iv

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Rabbit kidney proximal tubule cells in primary culture: Evaluation of the impact of expressed phenotype on cellular toxic response

Toxicology in Vitro ◽

10.1016/0887-2333(91)90058-l ◽

1991 ◽

Vol 5 (5-6) ◽

pp. 383-388 ◽

Cited By ~ 5

Author(s):

J.P. Morin ◽

C. Monteil ◽

R. Glomot ◽

J.P. Fillastre

Keyword(s):

Proximal Tubule ◽

Primary Culture ◽

Rabbit Kidney ◽

Toxic Response ◽

Proximal Tubule Cells ◽

Kidney Proximal Tubule ◽

Tubule Cells ◽

The Impact

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Increased functional differentiation of rabbit proximal tubule cells cultured in glucose-free media

AJP Renal Physiology ◽

10.1152/ajprenal.1992.263.1.f152 ◽

1992 ◽

Vol 263 (1) ◽

pp. F152-F162

Author(s):

A. Blais ◽

F. Jalal ◽

P. Crine ◽

J. Paiement ◽

A. Berteloot

Keyword(s):

Proximal Tubule ◽

Brush Border ◽

Brush Border Membrane ◽

Primary Cultures ◽

Morphological Differentiation ◽

Functional Expression ◽

Functional Differentiation ◽

Rabbit Kidney ◽

Proximal Tubule Cells ◽

Tubule Cells

We have determined the influence of glucose (Glc)-free medium on the growth and differentiation of rabbit kidney proximal tubule cells (PTC) in primary cultures. The specific growth rates and the protein-to-volume ratios are shown to be independent of the culture conditions. In contrast, the functional expression of four brush-border membrane enzyme markers was found to decline steadily and in the same way from day 3 in culture up to late confluence in Glc-containing medium, and different evolution patterns and high expression levels were observed up to confluence in a Glc-free glutamine (Gln)-supplemented medium. Electron microscopy clearly showed, however, that the functional and morphological differentiation of the brush-border membrane is not correlated. Finally, by use of an indirect immunofluorescent technique in combination with flow cytometry, it is demonstrated that confluent cells grown in Glc and Gln media form homogeneous cell populations of PTC. It is thus concluded that the functional differentiation of rabbit kidney PTC in primary cultures is strongly dependent upon the energy source present in the culture medium.

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