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Characterization and fabrication of nasal mucosal epithelial cell sheets derived from EGFP rats. (A) Schematic representation of the fabrication process for nasal mucosal epithelial cell sheets using EGFP rats. (B) Phase-contrast images 12 days after primary explant culture. (C) Phase-contrast micrographs of subcultured cells 7 days after seeding <t>onto</t> <t>temperature-responsive</t> culture dishes. (D) Macroscopic appearance of a cell sheet harvested by reducing the incubation temperature to 20 °C. (E) Colony-forming assay of cells harvested from the nasal mucosal epithelial cell sheet. (F) Fluorescence microscopy. Representative images of EGFP (green) and DAPI (blue) in the cell sheet. Scale bar = 50 μm. (G) Immunohistochemical staining for pan-cytokeratin. A representative cross-sectional image of the nasal mucosal epithelial cell sheet showing positive immunoreactivity for pan-cytokeratin (AE1/AE3). Scale bar = 25 μm.
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Characterization and fabrication of nasal mucosal epithelial cell sheets derived from EGFP rats. (A) Schematic representation of the fabrication process for nasal mucosal epithelial cell sheets using EGFP rats. (B) Phase-contrast images 12 days after primary explant culture. (C) Phase-contrast micrographs of subcultured cells 7 days after seeding <t>onto</t> <t>temperature-responsive</t> culture dishes. (D) Macroscopic appearance of a cell sheet harvested by reducing the incubation temperature to 20 °C. (E) Colony-forming assay of cells harvested from the nasal mucosal epithelial cell sheet. (F) Fluorescence microscopy. Representative images of EGFP (green) and DAPI (blue) in the cell sheet. Scale bar = 50 μm. (G) Immunohistochemical staining for pan-cytokeratin. A representative cross-sectional image of the nasal mucosal epithelial cell sheet showing positive immunoreactivity for pan-cytokeratin (AE1/AE3). Scale bar = 25 μm.
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Characterization and fabrication of nasal mucosal epithelial cell sheets derived from EGFP rats. (A) Schematic representation of the fabrication process for nasal mucosal epithelial cell sheets using EGFP rats. (B) Phase-contrast images 12 days after primary explant culture. (C) Phase-contrast micrographs of subcultured cells 7 days after seeding <t>onto</t> <t>temperature-responsive</t> culture dishes. (D) Macroscopic appearance of a cell sheet harvested by reducing the incubation temperature to 20 °C. (E) Colony-forming assay of cells harvested from the nasal mucosal epithelial cell sheet. (F) Fluorescence microscopy. Representative images of EGFP (green) and DAPI (blue) in the cell sheet. Scale bar = 50 μm. (G) Immunohistochemical staining for pan-cytokeratin. A representative cross-sectional image of the nasal mucosal epithelial cell sheet showing positive immunoreactivity for pan-cytokeratin (AE1/AE3). Scale bar = 25 μm.
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Characterization and fabrication of nasal mucosal epithelial cell sheets derived from EGFP rats. (A) Schematic representation of the fabrication process for nasal mucosal epithelial cell sheets using EGFP rats. (B) Phase-contrast images 12 days after primary explant culture. (C) Phase-contrast micrographs of subcultured cells 7 days after seeding <t>onto</t> <t>temperature-responsive</t> culture dishes. (D) Macroscopic appearance of a cell sheet harvested by reducing the incubation temperature to 20 °C. (E) Colony-forming assay of cells harvested from the nasal mucosal epithelial cell sheet. (F) Fluorescence microscopy. Representative images of EGFP (green) and DAPI (blue) in the cell sheet. Scale bar = 50 μm. (G) Immunohistochemical staining for pan-cytokeratin. A representative cross-sectional image of the nasal mucosal epithelial cell sheet showing positive immunoreactivity for pan-cytokeratin (AE1/AE3). Scale bar = 25 μm.
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Characterization and fabrication of nasal mucosal epithelial cell sheets derived from EGFP rats. (A) Schematic representation of the fabrication process for nasal mucosal epithelial cell sheets using EGFP rats. (B) Phase-contrast images 12 days after primary explant culture. (C) Phase-contrast micrographs of subcultured cells 7 days after seeding onto temperature-responsive culture dishes. (D) Macroscopic appearance of a cell sheet harvested by reducing the incubation temperature to 20 °C. (E) Colony-forming assay of cells harvested from the nasal mucosal epithelial cell sheet. (F) Fluorescence microscopy. Representative images of EGFP (green) and DAPI (blue) in the cell sheet. Scale bar = 50 μm. (G) Immunohistochemical staining for pan-cytokeratin. A representative cross-sectional image of the nasal mucosal epithelial cell sheet showing positive immunoreactivity for pan-cytokeratin (AE1/AE3). Scale bar = 25 μm.

Journal: Regenerative Therapy

Article Title: Characterization of the role and fate of nasal mucosal epithelial cell sheets in middle ear regeneration

doi: 10.1016/j.reth.2026.101130

Figure Lengend Snippet: Characterization and fabrication of nasal mucosal epithelial cell sheets derived from EGFP rats. (A) Schematic representation of the fabrication process for nasal mucosal epithelial cell sheets using EGFP rats. (B) Phase-contrast images 12 days after primary explant culture. (C) Phase-contrast micrographs of subcultured cells 7 days after seeding onto temperature-responsive culture dishes. (D) Macroscopic appearance of a cell sheet harvested by reducing the incubation temperature to 20 °C. (E) Colony-forming assay of cells harvested from the nasal mucosal epithelial cell sheet. (F) Fluorescence microscopy. Representative images of EGFP (green) and DAPI (blue) in the cell sheet. Scale bar = 50 μm. (G) Immunohistochemical staining for pan-cytokeratin. A representative cross-sectional image of the nasal mucosal epithelial cell sheet showing positive immunoreactivity for pan-cytokeratin (AE1/AE3). Scale bar = 25 μm.

Article Snippet: The cells were then seeded onto temperature-responsive cell culture inserts (CellSeed) at a density of 4 × 10 4 cells/cm 2 and cultured in KCM for 7 days.

Techniques: Derivative Assay, Incubation, Fluorescence, Microscopy, Immunohistochemical staining, Staining