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. 2014 Mar 14;11(7):2082–2091. doi: 10.1021/mp5000295

Figure 2.

Figure 2

Schematic figure showing different steps of fabrication and configuration of the 3D tissue engineered airway epithelium. Calu-3 epithelial cells are seeded onto one PET scaffold and MRC-5 fibroblasts are seeded onto a second, separate scaffold (A). Following 72 h culture, scaffolds are combined by layering the epithelial scaffold monolayer on top of the fibroblast scaffold layer to form the coculture model. Cells are subsequently cultured for 2 weeks at the ALI to allow for differentiation of the epithelial cells, including establishment of tight junctions (B). Monocyte-derived DCs are seeded onto separate PET scaffolds and then inserted into the coculture model. The upper epithelial scaffold is temporally lifted away from the lower MRC-5 fibroblast layer so that the separate third scaffold containing dendritic cells may be placed on-top of the MRC-5 scaffold layer (C). The Calu-3 layer is placed on-top of the DC scaffold layer, resulting in the DC layer sandwiched between the epithelial and fibroblast scaffold layers to form the triculture model (D).