Cell culture systems for isolation of SARS-CoV-2 clinical isolates and generation of recombinant virus

• Published in: iScience Vol. 26; no. 5; p. 106634
• Main Authors: Chen, Da-Yuan, Turcinovic, Jacquelyn, Feng, Shuchen, Kenney, Devin J., Chin, Chue Vin, Choudhary, Manish C., Conway, Hasahn L., Semaan, Marc, Close, Brianna J., Tavares, Alexander H., Seitz, Scott, Khan, Nazimuddin, Kapell, Sebastian, Crossland, Nicholas A., Li, Jonathan Z., Douam, Florian, Baker, Susan C., Connor, John H., Saeed, Mohsan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 19.05.2023; The Author(s); Elsevier
• Subjects: Methodology in biological sciences; Virology; Methodology in biological sciences; Virology
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2023.106634

A simple and robust cell culture system is essential for generating authentic SARS-CoV-2 stocks for evaluation of viral pathogenicity, screening of antiviral compounds, and preparation of inactivated vaccines. Evidence suggests that Vero E6, a cell line commonly used in the field to grow SARS-CoV-2, does not support efficient propagation of new viral variants and triggers rapid cell culture adaptation of the virus. We generated a panel of 17 human cell lines overexpressing SARS-CoV-2 entry factors and tested their ability to support viral infection. Two cell lines, Caco-2/AT and HuH-6/AT, demonstrated exceptional susceptibility, yielding highly concentrated virus stocks. Notably, these cell lines were more sensitive than Vero E6 cells in recovering SARS-CoV-2 from clinical specimens. Further, Caco-2/AT cells provided a robust platform for producing genetically reliable recombinant SARS-CoV-2 through a reverse genetics system. These cellular models are a valuable tool for the study of SARS-CoV-2 and its continuously emerging variants. [Display omitted] •Human cell lines overexpressing ACE2 and TMPRSS2 were tested for SARS-CoV-2 infection•Two cell lines, Caco-2 and HuH-6, exhibited remarkable susceptibility to infection•Caco-2 and HuH-6 cells allowed isolation of SARS-CoV-2 from clinical specimens•Caco-2 cells supported production of genetically stable recombinant SARS-CoV-2 Methodology in biological sciences; Virology