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Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells <t>(MUC5AC),</t> basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].
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Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells <t>(MUC5AC),</t> basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].
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Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells <t>(MUC5AC),</t> basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].
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Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells <t>(MUC5AC),</t> basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].
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Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells <t>(MUC5AC),</t> basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].
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Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells <t>(MUC5AC),</t> basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].
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Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells (MUC5AC), basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Generation of a human vascularized 3D airway model replicating native mucosal heterogeneity

doi: 10.3389/fbioe.2026.1761561

Figure Lengend Snippet: Mucociliary differentiation of tracheal airway epithelial cells and formation of vascular structures after 4 weeks of culture at air-liquid-interface. (A) Representative microscopic image of tri-culture stained with Periodic Acid-Schiff Reaction (PAS) - hematoxylin. (B) Representative immunofluorescent images of cell-type-specific markers, from top to bottom: Club cells (SCGB1A1), goblet cells (MUC5AC), basal cells (TP63 and KRT5), ciliated cells (Acetylated Tubulin), tight junctions (claudin 1) [adapted from ]. (C) Scanning electron microscopy images of ciliated surface of tri-culture. (D) Longitudinal section of multiciliated epithelial cell visualized by transmission electron microscopy. Structural parts of cilia indicated by abbreviations: ax = axoneme, bb = basal body, tz = transition zone, bf = basal foot. (E) Transversal section of multiple cilia demonstrating the central microtubule pair surrounded by 9 doublets (9 + 2 configuration). (F) Projection of a 3D stack made by two-photon laser scanning microscopy of a tri-culture fibrin hydrogel with endothelial vessels (PECAM1, red) and stromal cells (vimentin, green) [adapted from ].

Article Snippet: MUC5B , Glycoprotein mucin 5B present in both the epithelial secretory cells and the submucosal glands ( ) , Rabbit , Invitrogen, PA5-82342 , 1:500.

Techniques: Staining, Electron Microscopy, Transmission Assay, Laser-Scanning Microscopy