Isolation, culture, and preliminary characterization of mucin‐producing cells from trachea of the domestic fowl

Abstract

Primary cell cultures enriched in mucin-producing cells and basal cells were established from the trachea of the domestic fowl. Epithelial cells were selectively removed from the trachea after incubation in 0.1% pronase/0.1% EDTA in Moscona's saline. The majority of the ciliated cells were removed during the initial 30 minutes of incubation. After 50 minutes of incubation, aggregates of mucin-producing cells and basal cells were removed in large numbers. The cellular aggregates rapidly attached to a collagen-coated substratum and the cells spread out on the culture surface. The mucin-producing cells retained their AB/PAS-reactive secretory granules. The basal cells replicated and as the culture approached confluency, these cells developed a fine dusting of AB/PAS-reactive material; later, larger secretory granules appeared in the cells. These observations suggest that mucin-producing cells are capable of retaining their AB/PAS-reactive secretory products in primary culture and that basal cells are capable of differentiating into mucin-producing cells in vitro.

Contribution of in vitro culture methods for respiratory epithelial cells to the study of the physiology of the respiratory tract

“…This has been demonstrated for tracheal cells from hamsters [27][28][29][30][31], humans [21,22,24), rabbits [32][33][34], dogs (9,35], ferrets [36] and rats [37] and for human nasal cells (6]. Other animals investigated are guinea-pig and monkey [38], domestic fowl [39] and swine [40].…”

“…Although isolation and culture of specific cell types such as Clara cells [ 44] and basal and goblet cells (39,40,45] is possible, most cultures are started from nonselected surface respiratory epithelial cells. Recently, monolayer culture techniques for human tracheobronchial submucosal glands have been developed [46,47].…”

“…The use of high (10-20%) serum concentrations [27,30,39,40] has been abandoned because serum inhibits growth [21,22] and induces squamous differentiation [6,34,57] possibly due to the presence of transforming growth factor ~. [58,59].…”