CULTURE AND CHARACTERIZATION OF HUMAN NASAL GLAND-CELLS

To study the regulation of ion transport by human nasal gland (HNG) ce lls, we developed a system for culturing HNG cells isolated by enzymat ic digestion from human nasal polyps. When plated on flasks in media c ontaining Ultroser G serum substitute and a variety of growth factors, the HNG cells became confluent after 5-7 days. Cells were collected b y trypsinization and replated at 10(6) cells/cm(2) on porous filters. Confluent monolayers formed on days 3-5 after replating and were studi ed on days 5-7. Transepithelial resistance and short-circuit current ( I-sc) were 177 +/- 15 Omega . cm(2) and 12.2 +/- 1.3 mu A/cm(2) (means +/- SE, n = 35). Acetylcholine (ACh, 10(-5) M) induced a transient in crease in I-sc by 4.4 +/- 1.0 mu A/cm(2) when added to both apical and basolateral sides and returned to baseline levels within 5 min. Diphe nylamine -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 i nhibit ACh-induced I-sc responses. ACh (10(-5) M) also induced an incr ease 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 respo nse to cholinergic agent via an increase in intracellular Ca2+ concent ration.