Glandular‐like morphogenesis and secretory activity of human tracheal gland cells in a three‐dimensional collagen gel matrix

Abstract

The extracellular matrix has been demonstrated to affect the differentiation of epithelial cells. We present evidence that in a three-dimensional (3-D) type I collagen gel matrix, isolated human adult tracheal gland (HTG) cells are capable of reconstructing new functional gland-like tubules in vitro. During the first two weeks in culture, HTG cells developed globular epithelial cell aggregates in which lumina is absent. By the third week in culture, the tubulogenesis and the formation of branching structures became evident with a polarized morphology, which in many aspects resembles the in vivo morphology. A central lumen was lined by polarized secretory epithelial cells exhibiting well-developed microvilli and apical secretory granules. Furthermore, we showed that the capacity of in vitro tracheal gland differentiation was associated with the basal deposition of laminin and type IV collagen around the gland-like tubules. A cell-associated 72 kDa type IV collagenase was expressed in developing tubule cells, as shown by immunocytochemistry. The secretion of the antileucoprotease (ALP), a protein marker of tracheal gland serous cells, was bidirectional in gland-like tubules, since up to 65% of released ALP was in the basolateral direction. Taken together, these observations indicate that isolated HTG cells in a 3-D collagen matrix form functional tracheal gland-like tubules and suggest that similar new tracheobronchial gland formations may occur during the human normal gland development and remodeling.

Development of Glandular Models from Human Nasal Progenitor Cells

“…Tubules express MUC5B and lysozyme, but not acetylated a-tubulin or MUC5AC, and thus exhibited similar molecular, but not structural, characteristics with acini. A study in which human nasal polyp surface epithelial cells were grown on Col I for 13 days showed tubules with similar morphological structures to ours, with two different types of tubules observed by electron microscopy (20,21). In one type, there were numerous ciliated cells as well as preciliated cells that exhibit secretory granules and centrioles migrating toward the cell apex.…”

“…Matrigel also promotes the differentiation of HBE cells into glandular acini that express glandular markers (12). Col I promotes the organization of nasal polyp epithelial and HBE cells into tube-like structures (20,21). HBE cells cocultured with fetal lung fibroblasts on Col I form tubular structures that undergo dichotomous branching (22), implying that ECM and fibroblasts are associated with glandular development.…”

High Susceptibility for Cystic Fibrosis Human Airway Gland Cells to Produce IL-8 Through the IκB Kinase α Pathway in Response to Extracellular NaCl Content

“…Penicillin G (100 U/ml) and streptomycin (100 U/ml) were also added. After 4 wk in primary culture, second-and third-passage CF-HBG and non-CF HBG cells had proliferated and exhibited characteristics of homogenous submucosal epithelial and secretory gland cells, as previously described (18,19). Using the halide-sensitive fluorescent dye 6-methoxy-N-(3-sulfopropyl)-quinolinium, we have previously shown a significant increase in Cl Ϫ channel activity via the CFTR protein in non-CF HBG cells in response to forskolin treatment (demonstrating a cAMP-dependent activation of a Cl Ϫ efflux), which is not preserved in cultured CF HBG cells (7).…”

Intracellular free Ca2+ dynamic changes to histamine are reduced in cystic fibrosis human tracheal gland cells

“…Isolation and culture of CF-HTG and HTG cells were performed as previously described [10,22]. The CF-HTG cells were homozygote for the AF 508 CFTR deletion (AF 508/AF 508).…”

“…In brief, cells were isolated by enzymatic digestion from trachea mucosas and were grown onto type 1 collagen coated flasks in a Dulbecco's modified Eagle's medium/Ham's FI2 mixture (50/50%, v/ v) supplemented with 100 U/ml penicillin G and 100 lag/lal streptomycin as antibiotics and with 1% Ultroser G (IBF, Villeneuve la Garenne, France), 3 laM epinephrine, glucose and sodium pyruvate made up to 10 g/1 and 0.33 g/l, respectively. In these culture conditions, cells proliferate and exhibit characteristics of epithelial and serous secretory cell type [10,22].…”