Abstract
Virus propagation methods generally use transformed cell lines to grow viruses from clinical specimens, which may force viruses to rapidly adapt to cell culture conditions, a process facilitated by high viral mutation rates. Upon propagation in VeroE6 cells, SARS-CoV-2 may mutate or delete the multibasic cleavage site (MBCS) in the spike protein. Previously, we showed that the MBCS facilitates serine protease-mediated entry into human airway cells (Mykytyn et al., 2021). Here, we report that propagating SARS-CoV-2 on the human airway cell line Calu-3-that expresses serine proteases-prevents cell culture adaptations in the MBCS and directly adjacent to the MBCS (S686G). Similar results were obtained using a human airway organoid-based culture system for SARS-CoV-2 propagation. Thus, in-depth knowledge on the biology of a virus can be used to establish methods to prevent cell culture adaptation.
| Original language | English |
|---|---|
| Article number | e66815 |
| Journal | eLife |
| Volume | 10 |
| DOIs | |
| Publication status | Published - 9 Apr 2021 |
Bibliographical note
Funding Information:This work was supported by the Netherlands Organization for Health Research and
Funding Information:
Development (ZONMW) grant agreement 10150062010008 to B.L.H and co-funded by the
Funding Information:
This work was supported by the Netherlands Organization for Health Research and Development (ZONMW) grant agreement 10150062010008 to B.L.H and co-funded by the PPP Allowance (grant agreement LSHM19136) made available by Health Holland, Top Sector Life Sciences & Health, to stimulate public-private partnerships. The present manuscript was part of the research program of the Netherlands Centre for One Health. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
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