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
Affiliations
Da-Yuan Chen
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA
Jacquelyn Turcinovic
National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Department of Microbiology, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA
Shuchen Feng
Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine, Maywood, IL, USA
Devin J. Kenney
National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Department of Microbiology, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA
Chue Vin Chin
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA
Manish C. Choudhary
Brigham and Women’s Hospital, Boston, MA, USA; Harvard Medical School, Cambridge, MA, USA
Hasahn L. Conway
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA
Marc Semaan
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA
Brianna J. Close
National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Department of Microbiology, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA
Alexander H. Tavares
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA
Scott Seitz
National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Department of Microbiology, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA
Nazimuddin Khan
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA
Sebastian Kapell
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA
Nicholas A. Crossland
National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Department of Pathology and Laboratory Medicine, Boston University School of Medicine, Boston, MA, USA
Jonathan Z. Li
Brigham and Women’s Hospital, Boston, MA, USA; Harvard Medical School, Cambridge, MA, USA
Florian Douam
National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Department of Microbiology, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA
Susan C. Baker
Department of Microbiology and Immunology, Loyola University Chicago Stritch School of Medicine, Maywood, IL, USA
John H. Connor
National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Department of Microbiology, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA
Mohsan Saeed
Department of Biochemistry, Boston University Chobanian and Avedisian School of Medicine, Boston, MA, USA; National Emerging Infectious Diseases Laboratories, Boston University, Boston, MA, USA; Corresponding author
Summary: 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.
