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

Da-Yuan Chen; Jacquelyn Turcinovic; Shuchen Feng; Devin J Kenney; Chue Vin Chin; Manish C Choudhary; Hasahn L Conway; Marc Semaan; Brianna J Close; Alexander H Tavares; Scott Seitz; Nazimuddin Khan; Sebastian Kapell; Nicholas A Crossland; Jonathan Z Li; Florian Douam; Susan C Baker; John H Connor; Mohsan Saeed

doi:10.1016/j.isci.2023.106634

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

iScience. 2023 May 19;26(5):106634. doi: 10.1016/j.isci.2023.106634. Epub 2023 Apr 10.

Authors

Da-Yuan Chen^{1

2}, Jacquelyn Turcinovic^{2

3}, Shuchen Feng⁴, Devin J Kenney^{2

3}, Chue Vin Chin^{1

2}, Manish C Choudhary^{5

6}, Hasahn L Conway^{1

2}, Marc Semaan^{1

2}, Brianna J Close^{2

3}, Alexander H Tavares^{1

2}, Scott Seitz^{2

3}, Nazimuddin Khan^{1

2}, Sebastian Kapell^{1

2}, Nicholas A Crossland^{2

7}, Jonathan Z Li^{5

6}, Florian Douam^{2

3}, Susan C Baker⁴, John H Connor^{2

3}, Mohsan Saeed^{1

2}

Affiliations

¹ Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA.
² National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA.
³ Department of Microbiology, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA.
⁴ Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine, Maywood, IL, USA.
⁵ Brigham and Women's Hospital, Boston, MA, USA.
⁶ Harvard Medical School, Cambridge, MA, USA.
⁷ Department of Pathology and Laboratory Medicine, Boston University School of Medicine, Boston, MA, USA.

Abstract

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.

Keywords: Methodology in biological sciences; Virology.