Expression of Mucin Synthesis and Secretion in Human Tracheobronchial Epithelial Cells Grown in Culture

Abstract

The effects of culture conditions on growth and differentiation of human tracheobronchial epithelial (HTBE) cells have been defined. Epithelial cells were dissociated from tissues by protease treatment and were plated on tissue culture dishes in F12 medium supplemented with insulin, transferrin, epidermal growth factor, hydrocortisone, cholera toxin, bovine hypothalamus extract, and retinol. HTBE cells did not express any mucociliary function (ciliogenesis or mucin secretion) on tissue culture plastic, but they could be passaged 3 to 5 times with a total of 10 to 25 population doublings. Cells from early passages re-express both these functions when transplanted to tracheal grafts. When tissue culture plates were coated with collagen film or collagen gel substrata, cell attachment and proliferation were stimulated. However, the expression of mucous cell function in culture occurred only when cells were plated on collagen gel substrata and vitamin A (retinol) was present in the medium. Mucous cell differentiation under optimal conditions was defined by ultrastructural studies, by immunologic studies with mucin-specific monoclonal antibodies, and by carbohydrate and amino acid compositional analyses of mucin-like glycoproteins purified from culture medium. These results demonstrate for the first time that HTBE cells can express mucin synthesis and secretion under appropriate culture conditions.

Enhancement of Calu-1 human lung carcinoma cell growth in serum-free medium by retinoids: dependence on AP-1 activation, but not on retinoid response element activation

“…This was an unexpected ®nding because retinoids inhibit the growth of a variety of malignant cell lines (Amos and Lotan, 1991;Gudas et al, 1994;Lotan, 1995), and we showed that several other NSCLC cell lines were more sensitive to ATRA in serum-free than in serum-containing medium. Similar to our ®nding, the growth of normal human tracheobronchial epithelial cells (Lechner et al, 1982;Wu et al, 1990) and of human lung adenocarcinoma cells (Yang et al, 1992) was enhanced by low ATRA concentrations (510…”

“…yet inhibited at high concentrations (10 76 M) (Wu et al, 1990;Geradts et al, 1993). The growth of SCLC cell lines was inhibited by 13-cis-RA concentrations 410 76 M in serum-free medium but not in the presence of serum and 2% bovine serum albumin (BSA), which was identi®ed as the major serum component responsible for binding radiolabeled ATRA, reversed the growth inhibitory eect of 13-cis-RA (Avis et al, 1995).…”

IL-17 Markedly Up-Regulates β-Defensin-2 Expression in Human Airway Epithelium via JAK and NF-κB Signaling Pathways

“…Tissues were collected only from patients without known respiratory tract diseases. Primary cultures derived from these airway tissues have been established before (37,38). Protease-dissociated TBE cells were plated on a collagen gel substratum-coated Transwell (Corning Costar, Corning, NY) chamber (25 mm) at 1-2 ϫ 10 4 cells/ cm 2 , in a Ham's F12/DMEM (1:1) supplemented with insulin (5 g/ml), transferrin (5 g/ml), epidermal growth factor (10 ng/ml), dexamethasone (0.1 M), cholera toxin (10 ng/ml), bovine hypothalamus extract (15 g/ ml), BSA (0.5 mg/ml), and all-trans-retinoic acid (30 nM).…”

Potentiation of IL-19 expression in airway epithelia by IL-17A and IL-4/IL-13: Important implications in asthma

“…NHBE cells were maintained in air-liquid interface culture for full mucociliary differentiation. 20,21 The immortalized NHBE cell line HBE1 was cultured under the same conditions as NHBE cells. 19 Tracheobronchial tissue sections from patients with asthma, chronic obstructive pulmonary disease (COPD), and cystic fibrosis (CF) and from patients with no obvious airway diseases were obtained with consent from the University of Miami Medical Center.…”