Characterization of a mouse cortical collecting duct cell line.
A cortical collecting duct (CCD) cell line has been developed from a mouse transgenic for the early region of simian virus 40, Tg(SV40E)Bri/7. CCDs were microdissected and placed on collagen gels. Monolayers were subsequently subcultured onto permeable collagen membranes and maintained in serum-supplemented medium. One line, designated M-1, retained many characteristics of the CCD, including a typical epithelial appearance and CCD-specific antigens. M-1 cells, when grown in monolayers on permeable supports, exhibited a high transepithelial resistance (885.7 +/- 109.6 ohms/cm2) and developed a lumen negative transepithelial potential difference (PD) of -45.7 +/- 3.5 mV. The associated short-circuit current (SCC) averaged 71.8 +/- 10.3 microA/cm2, and was reduced by 95% by luminal application of amiloride. The cultured cells responded to arginine vasopressin (AVP) with a significant increase in SCC. M-1 cells generated significant transepithelial solute gradients. After 24 hours incubation, the composition of the luminal (L) and basolateral (B) media (in mM) was: [Na+], L = 106.7 +/- 0.9 and B = 127.4 +/- 0.4; [K+], L = 8.6 +/- 0.6 and B = 2.1 +/- 0.3; [Cl], L = 68.6 +/- 5.8 and B = 101.8 +/- 6.6; [HCO3], L = 15.5 +/- 1.5 and B = 8.6 +/- 1.2; while pH was 7.16 +/- 0.03 at the luminal and 6.94 +/- 0.03 at the basolateral side. The formation of these concentration gradients indicates that the CCD cultures absorb Na+ and Cl- and secrete K+.(ABSTRACT TRUNCATED AT 250 WORDS)
Stoos
B A
BA
Henry Ford Hospital, Hypertension Research Division, Detroit, Michigan.
Náray-Fejes-Tóth
A
A
Carretero
O A
OA
Ito
S
S
Fejes-Tóth
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D001692:Biological Transport; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D004594:Electrophysiology; D007150:Immunohistochemistry; D007477:Ions; D007672:Kidney Cortex; D007685:Kidney Tubules, Collecting / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D008855:Microscopy, Electron, Scanning
D000911:Antibodies, Monoclonal; D007477:Ions; D000242:Cyclic AMP