AJP - Renal Physiology, Vol 238, Issue 2 140-F149, Copyright © 1980 by American Physiological Society
Isolation of epithelial cells from toad bladder
H. J. Rodriguez, D. W. Scholer, M. L. Purkerson and S. Klahr
The epithelial cells of the toad bladder have been isolated by brief
exposure to ethylenediaminetetraacetic acid followed by treatment with
collagenase, DNAse, and the application of shearing forces. This approach
eliminates the need for scraping of the mucosal surface and reduces
mechanical damage during harvesting of the epithelium. The method yields
intact, isolated epithelial cells and few clumps. The three major types of
epithelial cells described in the intact toad bladder were present in the
final preparation. The cellular contents of nucleic acids and proteins (in
pg/cell) were: DNA 22.5 +/- 1.1; RNA, 12.9 +/- 0.6; and protein, 192 +/- 9.
The isolated cells possess rates of oxygen consumption and amino acid
incorporation higher than those of epithelial sheets obtained by
collagenase treatment and scraping of the intact bladder. However, the
production of cyclic nucleotides in response to stimulation by vasopressin
and carbachol is comparable in both preparations.
