Culture and characterization of human nasal gland cells

Abstract

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 10(6) 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 (Isc) were 177 +/- 15 omega.cm2 and 12.2 +/- 1.3 microA/cm2 (means +/- SE, n = 35). Acetylcholine (ACh, 10(-5) M) induced a transient increase in Isc by 4.4 +/- 1.0 microA/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 Isc by 30.2 +/- 1.7% (n = 6) and inhibited Isc responses induced by ACh. Amiloride also decreased baseline Isc by 12.3 +/- 3.4% (n = 6) but failed to inhibit ACh-induced Isc 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.