A cautionary perspective regarding the isolation and serial propagation of SARS-CoV-2 in Vero cells

doi:10.1038/s41541-021-00346-z

. 2021 Jun 17;6(1):83.

doi: 10.1038/s41541-021-00346-z.

A cautionary perspective regarding the isolation and serial propagation of SARS-CoV-2 in Vero cells

Simon G P Funnell^{1

2

3}, Babak Afrough¹, John James Baczenas⁴, Neil Berry⁵, Kevin R Bewley¹, Rebecca Bradford⁶, Clint Florence⁷, Yann Le Duff⁵, Mark Lewis⁷, Ryan V Moriarty⁴, Shelby L O Connor⁴, Karen L Osman¹, Steven Pullan¹, Sujatha Rashid⁶, Kevin S Richards^{1

8}, Kimberly J Stemple⁷, Ivana Knezevic⁹

Affiliations

¹ PHE Porton Down, National Infection Service, Public Health England, Salisbury, Wiltshire, UK.
² Quadram Institute Bioscience, Norwich Research Park, Norfolk, UK.
³ World Health Organization, Geneva, Switzerland.
⁴ Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, USA.
⁵ Division of Infectious Disease Diagnostics, National Institute for Biological Standards and Control, London, UK.
⁶ Biodefense and Emerging Infections Resources, NIH, Bethesda, MD, USA.
⁷ Office of Biodefense, Research Resources and Translational Research, Division of Microbiology and Infectious Diseases, NIAID, NIH, Bethesda, MD, USA.
⁸ Faculty of Health and Life Sciences, Oxford Brookes University, Oxford, UK.
⁹ World Health Organization, Geneva, Switzerland. knezevici@who.int.

PMID: 34140522
PMCID: PMC8211640
DOI: 10.1038/s41541-021-00346-z

A cautionary perspective regarding the isolation and serial propagation of SARS-CoV-2 in Vero cells

Simon G P Funnell et al. NPJ Vaccines. 2021.

. 2021 Jun 17;6(1):83.

doi: 10.1038/s41541-021-00346-z.

Authors

Affiliations

¹ PHE Porton Down, National Infection Service, Public Health England, Salisbury, Wiltshire, UK.
² Quadram Institute Bioscience, Norwich Research Park, Norfolk, UK.
³ World Health Organization, Geneva, Switzerland.
⁴ Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, USA.
⁵ Division of Infectious Disease Diagnostics, National Institute for Biological Standards and Control, London, UK.
⁶ Biodefense and Emerging Infections Resources, NIH, Bethesda, MD, USA.
⁷ Office of Biodefense, Research Resources and Translational Research, Division of Microbiology and Infectious Diseases, NIAID, NIH, Bethesda, MD, USA.
⁸ Faculty of Health and Life Sciences, Oxford Brookes University, Oxford, UK.
⁹ World Health Organization, Geneva, Switzerland. knezevici@who.int.

PMID: 34140522
PMCID: PMC8211640
DOI: 10.1038/s41541-021-00346-z

Abstract

An array of SARS-CoV-2 virus variants have been isolated, propagated and used in in vitro assays, in vivo animal studies and human clinical trials. Observations of working stocks of SARS-CoV-2 suggest that sequential propagation in Vero cells leads to critical changes in the region of the furin cleavage site, which significantly reduce the value of the working stock for critical research studies. Serially propagating SARS-CoV-2 in Vero E6 cells leads to rapid increases in genetic variants while propagation in other cell lines (e.g. Vero/hSLAM) appears to mitigate this risk thereby improving the overall genetic stability of working stocks. From these observations, investigators are urged to monitor genetic variants carefully when propagating SARS-CoV-2 in Vero cells.

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Conflict of interest statement

S.L.O. and J.J.B. manage a fee-for-service pathogen sequencing unit that is a part of Research Services at the Wisconsin National Primate Research Center. All other authors declare no competing interests.

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References

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1. Davidson AD, et al. Characterisation of the transcriptome and proteome of SARS-CoV-2 reveals a cell passage induced in-frame deletion of the furin-like cleavage site from the spike glycoprotein. Genome Med. 2020;12:68. doi: 10.1186/s13073-020-00763-0. - DOI - PMC - PubMed
1. Ogando NS, et al. SARS-coronavirus-2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology. J. Gen. Virol. 2020;101:925–940. doi: 10.1099/jgv.0.001453. - DOI - PMC - PubMed
1. Wilm A, et al. LoFreq: a sequence-quality aware, ultra-sensitive variant caller for uncovering cell-population heterogeneity from high-throughput sequencing datasets. Nucleic Acids Res. 2012;40:11189–11201. doi: 10.1093/nar/gks918. - DOI - PMC - PubMed
1. Caly L, et al. Isolation and rapid sharing of the 2019 novel coronavirus (SARS-CoV-2) from the first patient diagnosed with COVID-19 in Australia. Med. J. Aust. 2020;212:459–462. doi: 10.5694/mja2.50569. - DOI - PMC - PubMed

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[1] Hoffmann M, Kleine-Weber H, Pohlmann S. A multibasic cleavage site in the spike protein of SARS-CoV-2 is essential for infection of human lung cells. Mol. Cell. 2020;78:779.e5–784.e5. doi: 10.1016/j.molcel.2020.04.022. - DOI - PMC - PubMed

[2] Hoffmann M, Kleine-Weber H, Pohlmann S. A multibasic cleavage site in the spike protein of SARS-CoV-2 is essential for infection of human lung cells. Mol. Cell. 2020;78:779.e5–784.e5. doi: 10.1016/j.molcel.2020.04.022. - DOI - PMC - PubMed

[3] Davidson AD, et al. Characterisation of the transcriptome and proteome of SARS-CoV-2 reveals a cell passage induced in-frame deletion of the furin-like cleavage site from the spike glycoprotein. Genome Med. 2020;12:68. doi: 10.1186/s13073-020-00763-0. - DOI - PMC - PubMed

[4] Davidson AD, et al. Characterisation of the transcriptome and proteome of SARS-CoV-2 reveals a cell passage induced in-frame deletion of the furin-like cleavage site from the spike glycoprotein. Genome Med. 2020;12:68. doi: 10.1186/s13073-020-00763-0. - DOI - PMC - PubMed

[5] Ogando NS, et al. SARS-coronavirus-2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology. J. Gen. Virol. 2020;101:925–940. doi: 10.1099/jgv.0.001453. - DOI - PMC - PubMed

[6] Ogando NS, et al. SARS-coronavirus-2 replication in Vero E6 cells: replication kinetics, rapid adaptation and cytopathology. J. Gen. Virol. 2020;101:925–940. doi: 10.1099/jgv.0.001453. - DOI - PMC - PubMed

[7] Wilm A, et al. LoFreq: a sequence-quality aware, ultra-sensitive variant caller for uncovering cell-population heterogeneity from high-throughput sequencing datasets. Nucleic Acids Res. 2012;40:11189–11201. doi: 10.1093/nar/gks918. - DOI - PMC - PubMed

[8] Wilm A, et al. LoFreq: a sequence-quality aware, ultra-sensitive variant caller for uncovering cell-population heterogeneity from high-throughput sequencing datasets. Nucleic Acids Res. 2012;40:11189–11201. doi: 10.1093/nar/gks918. - DOI - PMC - PubMed

[9] Caly L, et al. Isolation and rapid sharing of the 2019 novel coronavirus (SARS-CoV-2) from the first patient diagnosed with COVID-19 in Australia. Med. J. Aust. 2020;212:459–462. doi: 10.5694/mja2.50569. - DOI - PMC - PubMed

[10] Caly L, et al. Isolation and rapid sharing of the 2019 novel coronavirus (SARS-CoV-2) from the first patient diagnosed with COVID-19 in Australia. Med. J. Aust. 2020;212:459–462. doi: 10.5694/mja2.50569. - DOI - PMC - PubMed

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A cautionary perspective regarding the isolation and serial propagation of SARS-CoV-2 in Vero cells

Affiliations

A cautionary perspective regarding the isolation and serial propagation of SARS-CoV-2 in Vero cells

Authors

Affiliations

Abstract

Conflict of interest statement

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Cited by

References

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