Immortalization and characterization of proximal tubule cells derived from kidneys of spontaneously hypertensive and normotensive rats.
Immortalization and characterization of proximal tubule cells derived from kidneys of spontaneously hypertensive and normotensive rats.
复制标题
自发性高血压和正常血压大鼠肾脏近端小管细胞的永生化和表征。
DOI：
10.1038/ki.1996.295
复制
发表时间：
1996
影响因子：
19.6
通讯作者：
Hopfer,U
中科院分区：
文献类型：
--
作者：
Woost,PG;Orosz,DE;Jin,W;Frisa,PS;Jacobberger,JW;Douglas,JG;Hopfer,U
Immortalization and characterization of proximal tubule cells derived from kidneys of spontaneously hypertensive and normotensive rats.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.
