A conditionally immortalized cell line from murine proximal tubule.
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 gamma-interferon (IFN-gamma)-inducible, temperature sensitive SV40 large T antigen gene (tsA58) and [2] mice of the 129/SvEv strain, the background from which most embryonic stem (ES) cells are derived. Under permissive conditions (33 degrees C and in the presence of IFN-gamma), 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 degrees C, without IFN-gamma), the cells undergo differentiation coinciding with the disappearance of the large T antigen. By electron microscopy, tsMPT cells are polarized and show microvilli at their apical surface. tsMPT cells express brush border enzymes gamma-glutamyl transpeptidase and carbonic anhydrase IV. They possess Na(+)-dependent transport systems for Pi, 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.
Loghman-Adham
M
M
Howard Hughes Medical Institute, Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, USA.
Rohrwasser
A
A
Helin
C
C
Zhang
S
S
Terreros
D
D
Inoue
I
I
Lalouel
J M
JM
D000808:Angiotensinogen / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis; D002256:Carbonic Anhydrases / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods*; D002461:Cell Line, Transformed; D000242:Cyclic AMP / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D007150:Immunohistochemistry; D017136:Ion Transport / Q000187:drug effects / Q000502:physiology; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D010281:Parathyroid Hormone / Q000494:pharmacology; D007703:Peptidyl-Dipeptidase A / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D012964:Sodium / Q000378:metabolism
D000952:Antigens, Polyomavirus Transforming; D010281:Parathyroid Hormone; D012333:RNA, Messenger; D000808:Angiotensinogen; D012964:Sodium; D000242:Cyclic AMP; D007703:Peptidyl-Dipeptidase A; D002256:Carbonic Anhydrases