We also compared the replication of isolates to the lab strain in models of the human respiratory tract and found differences in respiratory tract tropism.

We made stocks of these isolates in immortalised cell lines. NL63 isolates replicated in LLC-MK2 cells and this was boosted by ACE2 and TMPRSS2 over-expression. 229E isolates replicated well in Huh7 cells, with a modest increase in the isolates when we over-expressed TMPRSS2.

We found these isolates showed the highest genetic variability in their Spikes compared to lab-adapted reference strains.

We used human airway and alveolar epithelial cultures to recover 15 contemporary isolates of human coronaviruses NL63, 229E and OC43.

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