Human airway cells prevent SARS-CoV-2 multibasic cleavage site cell culture adaptation

Mart M. Lamers, Anna Z. Mykytyn, Tim I. Breugem, Yiquan Wang, Douglas C. Wu, Samra Riesebosch, Petra B. van den Doel, Debby Schipper, Theo Bestebroer, Nicholas C. Wu, Bart L. Haagmans

Research output: Contribution to journalArticlepeer-review


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 languageEnglish (US)
Article numbere66815
StatePublished - Apr 2021


  • COVID-19
  • Calu-3
  • airway organoids
  • serine proteases
  • multibasic cleavage site
  • cell culture adaptation
  • SARS-CoV-2
  • Cell culture adaptation
  • Airway organoids
  • Serine proteases
  • Multibasic cleavage site

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • General Neuroscience


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