Tubule formation by human surface respiratory epithelial cells cultured in a three-dimensional collagen lattice

Abstract

Human surface respiratory epithelial (HSRE) cells from nasal polyps have been cultured within collagen lattices in a serum-free defined medium. Cell growth observed over a period of 12 days showed a population doubling time of 36 h. Under these culture conditions, we observed a contraction of the lattices. Phase-contrast light microscopy and transmission electron microscopy demonstrated that the HSRE cells formed tubular ductlike structures. Lumens formed by HSRE cells were surrounded by cuboidal-shaped polarized cells with numerous ciliated cells, secretory cells, and undifferentiated cells. Epidermal growth factor (EGF) was observed to stimulate the tubule formation and the contraction of the lattices. Videomicroscopic observations and analysis of the ciliary beating frequency (CBF) demonstrated that the cilia were homogeneously distributed on the whole apical surface of the ciliated cells and that their movement was well coordinated, with a CBF similar to that observed in outgrowth cells from cultured human nasal and tracheal epithelia. Immunofluorescent staining of basement membrane components synthesized and secreted by cells revealed the presence of type III collagen around the tubules. Type IV collagen and laminin were present in the cytoplasm and at the periphery of the cells. The biotin-streptavidin-gold immunocytochemical technique with monoclonal anti-mucin antibody showed intracellular localization of mucins in secretory granules of the secretory cells. With the use of substrate gel electrophoresis polyacrylamide gels impregnated with gelatin, collagenase activity was detected in the conditioned medium of the cultured HSRE cells. These results suggest that both three-dimensional collagen gel and soluble factors such as EGF regulate tubule formation by HSRE cells. Moreover, the capacity of the epithelial cells to contract the gel suggests they may be involved in the wound healing process.

Quantifying the Early Immune Response and Adaptive Immune Response Kinetics in Mice Infected with Influenza A Virus

“…The death rates of infected target cells were estimated to be between 2.1 and 13.5 day Ϫ1 , higher than our estimate of 1.2 day Ϫ1 . The difference in the epithelial cell death rates could be due to the relatively high virus production value assigned in our study, which was chosen based on direct measurements from in vitro systems using cultured airway epithelial cells (6,27,57). Based on these estimates, we fixed virus production at 100 EID 50 ⅐ ml Ϫ1 ⅐ cell Ϫ1 ⅐ day Ϫ1 and estimated innate virus clearance (c V ) as 4.2 day Ϫ1 .…”

Expression of Matrix Metalloproteinase Gelatinases A and B by Cultured Epithelial Cells from Human Bronchial Explants

“…Constitutive production of major gelatinase B and minor gelatinase A by HBECs may contribute to the basal remodeling of the subepithelial basal lamina as well as to the epithelial tubular morphogenesis possibly involved in the remodeling of bronchial mucosa following injury and inflammation (44). Indeed, several in vitro studies have shown that cultures of epithelial cells from different origins including mammary glands (45) and respiratory epithelium may organize into tubular structures (46). Extracellular matrix-degrading proteinases such as 72-kDa gelatinase have been shown to regulate mammary epithelial function during involution (47).…”

Rapid intranasal delivery of chloramphenicol acetyltransferase in the active form to different brain regions as a model for enzyme therapy in the CNS

“…Additionally, type IV collagen is the one of the more abundant ECM proteins in nasal epithelium (Benali, Tournier et al, 1993), suggesting that MMP-9 would be an effective enzyme for increasing permeability there. Our results showed that the 75 kDa CAT was transported into all the regions of brain within 15 min.…”