title: Conditional immortalization of bicarbonate-secreting intercalated cells from rabbit.
abstract: We have derived an immortalized cell line from primary cultures of bicarbonate-secreting intercalated cells from rabbit. Cells were transfected with a plasmid encoding a temperature-sensitive large T antigen of SV40 plus the neomycin resistance gene under the control of an SV40 promoter. Transfectants were selected for resistance to G418. One stably transfected clone, designated IC250, was subcloned to ensure clonality, and a subclone (clone C) was characterized in detail. The cells divide continuously at permissive temperature. At restrictive temperature, they cease dividing and assume morphological and transport properties of true bicarbonate-secreting intercalated cells. They express appropriate ultrastructural features, bind peanut lectin in an apical pattern, are rich in carbonic anhydrase, stain for proton-adenosinetriphosphatase in a basolateral pattern, and do not stain with antibodies to erythrocyte band 3. Most monolayers of transformed type B intercalated cells do not achieve a significant transepithelial resistance; those monolayers that are sufficiently electrically tight for electrophysiological studies are capable of chloride-dependent bicarbonate transport.
authors.affiliation: Department of Medicine, Washington University School of Medicine, St. Louis, Missouri.
authors.forename: J C
authors.initials: JC
authors.lastname: Edwards
authors.forename: J
authors.initials: J
authors.lastname: van Adelsberg
authors.forename: M
authors.initials: M
authors.lastname: Rater
authors.forename: D
authors.initials: D
authors.lastname: Herzlinger
authors.forename: J
authors.initials: J
authors.lastname: Lebowitz
authors.forename: Q
authors.initials: Q
authors.lastname: al-Awqati
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001639:Bicarbonates / Q000378:metabolism*; D002455:Cell Division; D002461:Cell Line, Transformed; D002712:Chlorides / Q000502:physiology; D007668:Kidney / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D011817:Rabbits; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
chemical_list: D001639:Bicarbonates; D002712:Chlorides; D000251:Adenosine Triphosphatases