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

Figure 4

Influenza virus sialic acid receptor detection on 2D and 3D ALI cultures of HNTEC. (a) Influenza virus sialic acid receptor detection on 2D cultures of HNTEC. HNTEC were seeded in 12-well plates cultured at an appropriate density, then fixed and permeabilized as described above, and labeled with the biotinylated lectins SNA, MAA-I, and MAA-II and PBS (as a blank control), respectively. These three lectin markers were measured by immunofluorescence protocols. The cellular nuclei were stained with DAPI. The lectins were stained with fluorescein avidin DCS. Magnification 40x. Scale bar, 50 μm. (b) Influenza virus sialic acid receptor detection on 3D ALI cultures of HNTEC. ALI 3D cultures of HNTEC were fixed, permeabilized, and labeled with the biotinylated lectins SNA, MAA-I, and MAA-II, respectively. The nuclei were stained with DAPI. The lectins were stained with fluorescein avidin DCS. Magnification 40x. Scale bar, 50 μm. (c) Expression of ACE2 mRNA in HNTEC and HNBEC. Total cellular RNAs were extracted, and real-time RT-PCR was performed to detect the expression of ACE2 mRNA. The cancer cell line A549 was used as control cells.

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