A conditionally immortalized cell line from murine proximal tubule

Abstract

We have developed a conditionally immortalized murine cell line with proximal tubule characteristics (tsMPT) and a background suitable for genetic manipulations. tsMPT was derived from the F1 progeny of crosses between: [1] a transgenic mouse harboring a γ-interferon (IFN-γ)-inducible, temperature sensitive SV40 large T antigen gene (tsA58) trod [2] mice of the 129/SvEv strain, the background from which most embryonic stem (ES) cells are derived. Under permissive conditions (33°C and in the presence of IFN-γ), tsMPT cells grow rapidly as monolayers with a doubling time of 23 hours; the large T antigen can be detected by immunocytochemistry and by Western blotting. When transferred to non-permissive conditions (39°C, without IFN-γ), the cells undergo differentiation coinciding with the disappearance of the large T antigen. By electron microscopy, tsM PT cells are polarized and show microvilli at their apical surface, tsMPT cells express brush border enzymes γ-glutamyl transpeptidase and carbonic anhydrase IV. They possess Na+- dependent transport systems for P(i), D-glucose and L-proline as well as an amiloride-insensitive Na+-H+ exchanger. Intracellular cAMP generation is stimulated by parathyroid hormone but not by arginine vasopressin. Angiotensinogen mRNA and protein are present in tsMPT with markedly higher Levels at non-permissive conditions. tsMPT cells should be a useful model for investigation of the functional features of the proximal tubule epithelium in relation to cellular differentiation.