Abstract
We have established an in vitro 3D system which recapitulates the human tracheo-bronchial mucosa comprehensive of the pseudostratified epithelium and the underlying stromal tissue. In particular, we reported that the mature model, entirely constituted of primary cells of human origin, develops key markers proper of the native tissue such as the mucociliary differentiation of the epithelial sheet and the formation of the basement membrane. The infection of the pseudo-tissue with a strain of NonTypeable Haemophilus influenzae results in bacteria association and crossing of the mucus layer leading to an apparent targeting of the stromal space where they release large amounts of vesicles and form macro-structures. In summary, we propose our in vitro model as a reliable and potentially customizable system to study mid/long term host-pathogen processes.
Highlights
In the last decades, the exploitation of transformed and/or immortalized cell line monocultures turned out to be a powerful approach to unravel the mechanisms of infection for various microbial pathogens, but to allow the characterization of thousands of drug compounds
We observed that the thickness of the formed mucus layer could be influenced by culture conditions, such as air-liquid interface (ALI) phase duration, stromal cellular density and composition and/or washing frequency of the apical surface to simulate the physiological displacement of mucus substances
The BEM shows a well-defined line of nuclei just above the collagen sheet suggesting the formation of a basal layer typical of the human pluristratified mucosal epithelium
Summary
The exploitation of transformed and/or immortalized cell line monocultures turned out to be a powerful approach to unravel the mechanisms of infection for various microbial pathogens, but to allow the characterization of thousands of drug compounds Despite their proved utility, the need for more accurate and physiological systems has driven researchers to develop models based on two or more cellular lineages, including epithelial and immune cells [1]. Supporting scaffolds and biomaterials provide the framework in which cells can deposit extracellular matrix components and differentiate to form a functionally relevant tissue In this context the use of different cellular types allows a nearly exact reproduction of human specific anatomical districts that are of exceptional value if one wants to follow host-pathogen interaction phenomena [2, 3]. The complexity of these systems allows the characterization of basic interactions such as bacterial
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