AJP - Regulatory, Integrative and Comparative Physiology, Vol 258, Issue 6 1355-R1362, Copyright © 1990 by American Physiological Society
Pharmacological responsiveness of dissociated native and cultured eccrine secretory coil cells
H. Yokozeki, K. Saga, F. Sato and K. Sato
Marshall Dermatology Research Laboratories, Department of Dermatology, University of Iowa College of Medicine, Iowa City 52242.
Methacholine (MCh)- and isoproterenol (Iso)-stimulated 14CO2 production was
compared between freshly dissociated rhesus sweat secretory coil cells
(mainly clear cells) and cultured cells (grown on a collagen-coated plastic
plate) derived from native cells. 14CO2 production was enhanced by MCh and
by Iso in native coil cells (but not in ductal cells) in a
pharmacologically specific and dose-dependent manner. 14CO2 production in
subcultured coil cells (19-45 days in culture) was only one-third to
one-fifth that of native cells. MCh-stimulated 14CO2 production was
inhibited by ouabain and furosemide in both native and cultured coil cells.
A decrease in 14CO2 production, of about one-half, was already evident in
primary cells cultured for less than 1 wk. The decreased pharmacological
responsiveness of the cultured coil cells was seen, although the cultured
cells showed the typical epithelioid appearance, abundant mitochondria, the
occasional presence of intercellular lacunae resembling intercellular
canaliculi, and the persistence of immunoreactive keratin. We conclude that
1) a primary culture of sweat gland cells can be initiated from dissociated
cells; 2) cultured sweat secretory coil cells qualitatively, but not
quantitatively, retain the pharmacological responsiveness and transport
activity of the native cells as determined by 14CO2 production; 3)
collagenase-dissociated cells represent an excellent in vitro system for
the study of glandular function at the cellular level; and 4) the decrease
in pharmacological responsiveness is not simply due to trypsin treatment
during harvesting of cultured cells, because that of organ-cultured,
intact, secretory coils also declines with time of culture.
