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Initiation and characterization of primary mouse kidney epithelial cultures

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Summary

Primary cultures of murine renal epithelial cells were established from a preparation of proximal tubule fragments. Confluent cultures exhibited multiple dome formation, indicating the presence of tight junctions and an intact transcellular transport process. Ultrastructural analysis revealed a monolayer of polarized cells, with a sparse but clearly defined microvillar surface facing the growth medium and a basolateral surface attached to the substratum. Cultures grown on collagen gels did not show domes. The epithelial monolayer exhibited several differentiated functions of the proximal tubule: a) parathyroid hormone (PTH)-stimulated cAMP synthesis; b) production of 24,25-dihydroxyvitamin D3 from 25-hydroxyvitamin D3; c) high alkaline phosphatase activity; and d) Na+-dependent transport of phosphate (Pi) and α-methylglucoside (α-MG). The sugar uptake was selectively inhibited by phlorizin, a competitive inhibitor of glucose uptake at the luminal membrane. Kinetic analysis revealed independent transport systems for Pi and α-MG, with Km values corresponding to the high affinity systems identified in brush border membrane vesicles derived from the proximal tubule. Pi uptake by the epithelial monolayers was regulated by the concentration of Pi in the growth medium. Phorbol esters and PTH did not exert an effect on Pi and α-MG transport in mouse primary cultures. The present study demonstrates that primary cultures provide a useful in vitro preparation to investigate renal proximal tubular function.

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Authors and Affiliations

  1. MRC Genetics Group, McGill University-Montreal Children's Hospital Research Institute, H3H 1P3, Montreal, Quebec, Canada

    C. L. Bell, H. S. Tenenhouse & C. R. Scriver

  2. Department of Biology, McGill University, H3H 1P3, Montreal, Quebec, Canada

    C. L. Bell, H. S. Tenenhouse & C. R. Scriver

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  1. C. L. Bell
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  2. H. S. Tenenhouse
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Cindy Bell was the recipient of an MRC Studentship Award. This work was supported by the MRC (Group in Medical Genetics).

This is publication number 88011 of the McGill University-Montreal Children's Hospital Research Institute.

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Bell, C.L., Tenenhouse, H.S. & Scriver, C.R. Initiation and characterization of primary mouse kidney epithelial cultures. In Vitro Cell Dev Biol 24, 683–695 (1988). https://doi.org/10.1007/BF02623606

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