title: Immortalization and characterization of proximal tubule cells derived from kidneys of spontaneously hypertensive and normotensive rats.
abstract: Epithelial cell lines from the proximal tubule of SHR and WKY rats were generated by microdissection, cell growth on 3T3 cell feeder layers, and transduction of the SV40 large T-antigen gene. The cell lines that formed confluent, electrically-resistive monolayers (basal conductance 1 to 20 mS/cm2) were selected for further study. Of these, cell lines generated from one rat did not show evidence of T-antigen expression or integration, and apparently immortalized spontaneously. Cell lines from three other rats expressed high levels of T-antigen, and showed evidence of integration of one or more copies of T-antigen. All cell lines formed polarized monolayers with apical microvilli, tight junctional complexes, and convolutions of the basolateral plasma membrane. Most cell lines grew in the absence of extracellular glucose indicating a capacity for gluconeogenesis. Sodium succinate cotransport and P2-purinergic receptor mediated signaling were demonstrated in all lines tested. The cell lines also showed that Na/H exchanger activity is regulated by angiotensin II. The results indicate that these cell lines express a proximal tubular phenotype, and are morphologically and functionally similar to primary cultures. These rat cell lines represent a new, potentially useful cell model for elucidating the cellular and molecular mechanisms of genetic differences in proximal tubule Na+ reabsorption.
authors.affiliation: Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio, USA.
authors.forename: P G
authors.initials: PG
authors.lastname: Woost
authors.forename: D E
authors.initials: DE
authors.lastname: Orosz
authors.forename: W
authors.initials: W
authors.lastname: Jin
authors.forename: P S
authors.initials: PS
authors.lastname: Frisa
authors.forename: J W
authors.initials: JW
authors.lastname: Jacobberger
authors.forename: J G
authors.initials: JG
authors.lastname: Douglas
authors.forename: U
authors.initials: U
authors.lastname: Hopfer
mesh_terms: D016475:3T3 Cells; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D006973:Hypertension / Q000378:metabolism*; D017136:Ion Transport; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D051379:Mice; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D015398:Signal Transduction; D012964:Sodium / Q000378:metabolism
chemical_list: D000804:Angiotensin II; D012964:Sodium