Yyculture and characterization of human nasal gland cells.
Furukawa, Masayuki, Mutsuo Yamaya, Katsuhisa Ikeda, Takeshi Oshima,
Tomoko Suzuki, Hidetada Sasaki, and Tomonori Takasaka.
Departments of Otorhinolaryngology and Geriatric Medicine, Tohoku
University School of Medicine, Sendai, Japan
APStracts 3:0081L, 1996.
To study the regulation of ion transport by the human nasal gland
(HNG) cells, we developed a system for culturing HNG cells isolated
by enzymatic digestion from human nasal polyps. When plated on flasks
in media containing Ultroser G serum substitute and a variety of
growth factors, the HNG cells became confluent after 5-7 days. Cells
were collected by tripsinization and replated at 106 cells[tilde]ncm
-2 on porous filters. Confluent monolayers formed on day 3-5 after
replating and were studied on day 5-7. Transepithelial resistance and
short-circuit current (Isc) were 177 [umlaut]u} 15 [circumflex]a
[tilde]ncm2 and 12.2 [umlaut]u} 1.3 mA[tilde]ncm-2 (means [umlaut]u}
SE, n=35). Acetylcholine (ACh, 10-5 M) induced transient increase in
Isc by 4.4 [umlaut]u} 1.0 mA[tilde]ncm-2 when added to both apical
and basolateral sides, and returned to baseline levels within 5 min.
Diphenylamine-2 carboxylate (10-3 M) decreased baseline Isc by 30.2
[umlaut]u} 1.7 % (n=6), and inhibited Isc responses induced by ACh.
Amiloride also decreased baseline Isc by 12.3 [umlaut]u} 3.4 % (n=6),
but failed to inhibit ACh-induced Isc responses. ACh (10-5 M) also
induced increase in intracellular Ca2+ concentration measured by
fura-2 from 60.5 [umlaut]u} 16.3 nM to 248.3 [umlaut]u} 45.3 nM
(n=4). These findings suggest that gland cells from human nasal
polyps can be grown in culture to produce epithelial cell sheets of
high resistance, which secrete Cl- in response to cholinergic agent
via an increase in intracellular Ca2+ concentration.
Received 5 January 1996; accepted in final form 15 May 1996.
APS Manuscript Number L7-6.
Article publication pending Am. J. Physiol. (Lung Cell. Mol.
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 5 June 96