Primary cultures of rabbit, bicarbonate-secreting, intercalated cells were isolated. Cells were transfected by electroporation with the pZipSVtsA58 plasmid encoding a temperature-sensitive large T antigen of SV40 plus the neomycin resistance gene under the control of an SV40 promoter. Following transfection, a line of G418 resistant cells was obtained and designated IC250. These cells were subcloned (Clone C) and characterized. The cells divide continuously at the permissive temperature of 32C. At the restrictive temperature of 40C, they cease dividing and assume morphological and transport properties of
true bicarbonate-secreting intercalated epithelia cells. They express appropriate ultrastructural features, bind peanut lectin in an apical pattern, are rich in carbonic anhydrase and stain for proton-adenosinetriphosphatase in a basolateral pattern. These cells reverse the polarity of several proteins and show different cytoskeltal organization depending on the seeding density. The cell line may be used to study terminal differentiation of intercalated cells. |
