Coupled CRC 2D and ALI 3D Cultures Express Receptors of Emerging Viruses and Are More Suitable for the Study of Viral Infections Compared to Conventional Cell Lines
H1N1 replication in 2D airway normal epithelial cells. (a) Expression of influenza A virus nucleoprotein in HNTEC and HNBEC. Cells were seeded at per well in 6-well plates 24 h before inoculation of the H1N1pdm virus. The 2D monolayer cultured cells were inoculated with viruses at multiplicity of infection (MOI) of 0.001. Infected cells were harvested at the indicated time points. Then, cells were fixed with 4% () paraformaldehyde and permeabilized with Triton X-100 and then labeled with the antibody against anti-influenza A virus nucleoprotein. Expression of influenza A virus nucleoprotein was detected by the immunofluorescence assay. The nuclear staining was used with 0.5 μg/ml DAPI. Magnification 20x. Scale bar, 100 μm. (b) Expression of influenza A virus nucleoprotein in A549 and 16HBE cells. (c) Quantitation of influenza A virus nucleoprotein-positive cells. The experiment was repeated for three times. Averaged data are . Differences between cells were assessed using the unpaired -test (;,, and ).