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. 2019 May 21;9:165. doi: 10.3389/fcimb.2019.00165

Figure 1.

Figure 1

Nasal tissue processing, immortalization, and model generation. (A–C) Processing primary nasal tissue for immortalization. (D–F) Nasal cell outgrowth from primary tissue (observed at top of frame) used for immortalization (20X magnification). (G,H) staining of cross sections of LLI cultures of immortalized NEC revealed a lack of differentiation and monolayer formation (H) (20X magnification). The transwell membrane can be seen beneath the cells. ALI culture of immortalized NEC led to differentiation and multilayer formation, closely resembling native tissue architecture. These studies were completed with similar results across 4 distinct NEC. (I–K) The apical surface of differentiated NEC visualized by SEM revealed microvilli and mucin vacuoles. (L) TEER values peaked by 7 days, indicating epithelial barrier integrity as shown by average values across the NEC cultures (n = 3 replicates for each NEC culture). SEM magnification is shown at 2 k (I), 15 k (J), and 25 k (K).