Isolation and Characterization of Primary Human Nasal Submucosal Gland Cell Culture

Kim, Woo Jeong; Park, Ji Hoon; Han, Seung Hoon; Lee, Sang Hag

Abstract

BACKGROUND: All tissue components in the nasal mucosa, including the epithelium, goblet cells, excretory glands, and blood capillaries participate in nasal secretory response. In vivo studies have suggested that, of these components, submucosal glands are thought to play an important role in nasal mucus secretion. These studies, however, are limited in dealing with pure secretory activity of submucosal glands. Therefore, in vitro study using cultured nasal submucosal glands is required to evaluate the pathophysiological mechanisms of nasal mucus secretion. The present study draws on experience obtained from the culture of human submucosal glands isolated from nasal mucosa.
MATERIALS AND METHODS
To obtain isolated submucosal gland cells, the uncinate process mucosa was stripped of surface epithelum by stroking the luminal surface with cotton tip applicator and minced into small fragments. By enzymatic treatment, thereafter, the submucosal gland cells were isolated and plated on culture flasks containing media. Using millipore inserts and cover slide, culuted cell was stained AB-PAS to identify that confluent monolayer cells were submucosal gland cell.
RESULTS
The uncinate process mucosa were appropriate for the isolation of submucosal gland cell. An average of 1.2X106 cells were obtained in every preparation of cells. The exclusion ratio of trypan blue was 95%. The isolated cells were strongly stained in AB-PAS, suggesting that cells are of glandular origin. Plated gland cell formed colony on the 3rd day and became confluent after 5-7 days. The proportion of the stained cells to the total cells decreased depending the duration of cell culture.
CONCLUSION
We succeeded in isolation and culture of submucosal gland cells which can provide model systems for studying the mechanisms of nasal glandular cell secretion in vivo.