Fig. 5
From: Long-term culture of chicken tracheal organoids for the purpose of avian influenza virus research
Replication of AIVs in CTO-derived 2D cultures as monoculture and co-culture with endothelial cells. CTO-derived 2D cultures (passage 7) cultured without (A–C) or with (D–F) chicken aortic endothelial cells on the basolateral side, were inoculated apically at 14 days post-seeding in duplicate with H7N7 LPAIV (HPAIV ΔMBCS) or HPAIV A/chicken/Netherlands/1/03 at an MOI of 0.0001. Detection of infectious virus titers in apical washes (A, D) and the basolateral medium (B, E) as determined by end-point titration assay in MDCK cells and expressed as log10 TCID50/mL. Each line represents a technical replicate. C, F Hematoxylin and eosin (H&E) and influenza virus nucleoprotein (NP, in dark red) stainings with hematoxylin counterstain of representative parts of the CTO-derived 2D cultures, fixed at 48 h post-inoculation and processed as formalin-fixed paraffin-embedded tissue. Created with BioRender.com