To study the regulation of ion transport by 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 trypsinization and replated at 106 cells/cm2 on porous filters. Confluent monolayers formed on days 3-5 after replating and were studied on days 5-7. Transepithelial resistance and short-circuit current (I(sc)) were 177 ± 15 Ω · cm2 and 12.2 ± 1.3 μA/cm2 (means ± SE, n = 35). Acetylcholine (ACh, 10-5 M) induced a transient increase in I(sc) by 4.4 ± 1.0 μA/cm2 when added to both apical and basolateral sides and returned to baseline levels within 5 min. Diphenylamine-2-carboxylate (10-3 M) decreased baseline I(sc) by 30.2 ± 1.7% (n = 6) and inhibited I(sc) responses induced by ACh. Amiloride also decreased baseline I(sc) by 12.3 ± 3.4% (n = 6) but failed to inhibit ACh- induced I(sc) responses. ACh (10-5 M) also induced an increase in intracellular Ca2+ concentration measured by fura 2-AM from 60.5 ± 16.3 to 248.3 ± 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.
|ジャーナル||American Journal of Physiology - Lung Cellular and Molecular Physiology|
|出版ステータス||Published - 1996 10|
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