Air–Liquid Interface Culture of Nasal Epithelial Cells on Denuded Amniotic Membranes

Abstract

Laboratory models of the respiratory lining with mucin secreting epithelial goblet cells have been obtained by culturing epithelial cells under air-liquid interface (ALI) conditions, which require specific wells with semi-permeable substrates. In this study nasal epithelial cells (NEC) were successfully cultured on denuded amniotic membrane (AM) under ALI conditions. The cells adhered well on both sides of the denuded AM (i.e., the basement membrane or spongy layer) and proliferated to confluency at the same time as cells grown on a synthetic membrane. The cytoskeleton structure of cells grown on denuded AMs appeared to be denser and firmer than that of cells cultured on synthetic membranes. Cultures on the denuded AM differentiated to contain goblet cells which produce and secrete mucins and ciliated cells. Cells cultured on the denuded AM were more stable under airflow conditions than cells on the synthetic membranes. The results of this study suggest that NEC culture on denuded AM and under ALI conditions creates a stable and well-differentiated in vitro model of the nasal lining for laboratory studies.

Improved In Vitro Model for Intranasal Mucosal Drug Delivery: Primary Olfactory and Respiratory Epithelial Cells Compared with the Permanent Nasal Cell Line RPMI 2650

“…Nevertheless, in vitro cell systems can be seen as simplified models of the first epithelial barrier. They can be used to study specific transporters for high molecular weight molecules and unspecific diffusion of small molecules [59,60,61]. Nevertheless, considering transport mechanism and cell type specific characteristic it is necessary to set stringent criteria for these cellular models.…”

Fluid-Flow Induced Wall Shear Stress and Epithelial Ovarian Cancer Peritoneal Spreading

“…For the flow experiments we cultured the EOC cells on the denuded AM, which was harvested from term placentas and denuded from the layer of epithelial cells, as described in an earlier study [15]. The denuded AM is a thick extra-cellular membrane made of an avascular stroma that contains collagen and fibronectin.…”

Preliminary Study on the Development of In Vitro Human Respiratory Epithelium Using Collagen Type I Scaffold as a Potential Model for Future Tracheal Tissue Engineering

“…However, in the classical submerged single-layer culture model, ciliary differentiation of epithelial cells did not occur [56]. The ALI cell culture model enabled the cells to undergo mucociliary differentiation [57]; this is of more importance regarding therapeutic application to enable the stem cells or progenitor cells to differentiate into the correct phenotype. The co-orientation of differentiated cells into a functional assembly of tissues is another vital prerequisite for successful and functional engineered tissue [58].…”