3D Reconstruction of the Human Airway Mucosa In Vitro as an Experimental Model to Study NTHi Infections

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.

Bioengineering the microanatomy of human skin

“…; Marrazzo et al. ). However, the use of primary cells directly from patients in our previous work demonstrated the issue of donor‐to‐donor variation (Hill et al.…”

“…The Alvetex â scaffold used in this study provides an inert and stable physical 3D microenvironment that enables cells to more closely recapitulate their native morphology (Maltman & Przyborski, 2010). Porous polystyrene scaffolds have previously been used successfully to develop complex multicellular tissue structures (Hill et al 2015;Marrazzo et al 2016). However, the use of primary cells directly from patients in our previous work demonstrated the issue of donor-to-donor variation (Hill et al 2015), further highlighting the need for cells to undergo quality control prior to their use in skin models, which can be achieved through the use of commercially available cells.…”

Animal Models Reflecting Chronic Obstructive Pulmonary Disease and Related Respiratory Disorders: Translating Pre-Clinical Data into Clinical Relevance

“…Primary cells obtained directly from patients through lung resections, bronchoalveolar lavage fluid extraction, and biopsies offer a more patient-specific phenotype compared to immortalized cell lines [142,143]. The utility of these cell lines is increased when used as structurally representative models such as air-liquid interface (ALI) cultures, which facilitate cell differentiation and the development of structural features such as cilia [144][145][146]. A noteworthy drawback of using in vitro cultured cells is their removal from the extracellular matrix and milieu of inflammatory cells, potentially skewing readouts and resulting in poor translational value for clinical application [143].…”

3D organ models—Revolution in pharmacological research?

“…They are typically generated by sequential cell seeding into cell culture inserts or porous 3D scaffolds. Of these, bone and cartilage [19], and epithelia such as skin [20,21], or lung [22] are currently the most advanced, while models of more complex tissues such as the brain are at earlier stages of development. Notably, organ models may vary in their complexity and, hence, predictivity.…”