Figure 6
SP GFP
+ derived cells form a complete epithelium in organotypic culture upon serial passage.
Samples were harvested with trypsin after formation of the primary epithelium (described in Figures 4 and 5) and harvested again after the initial organotypic culture for the second organotypic culture. (A) There were only a few scattered epithelial cells in serial samples prepared form the organotypic cultures of unsorted WT (GFP–) cells (H&E staining). (B–F) Immunohistochemistry revealed that the cells were GFP– (B), Ki67– (C), CK14– (D), CK4– (E), and CK13– (F). (G) Only a few isolated clusters of epithelial cells did not form an epithelium from 103 NSP cells from GFP+ mice mixed with 3 × 104 unsorted GFP– cells (H&E staining). (H–L) Immunohistochemistry revealed that the cells were mostly GFP– (H), Ki67– (I), minimally and irregularly CK14+ (J) and CK4+ (K), and CK13+ (L). (M) A complete epithelium formed, derived from sorted 103 SP cells from GFP+ mice mixed with 3 × 104 unsorted GFP– cells (H&E staining). (N–R) Immunohistochemistry revealed that cells were GFP+ (N), basal cells were Ki67+ (O) and CK14+ (P), and suprabasal cells were CK4+ (Q) and CK13+ (R). Dashed line represents basement membrane. Original magnification, ×100 (insets in A, G, and M); ×400 (B–F, H–L, and N–R). Scale bars: 25 μm.