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

iScience (May 2023)

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

Journal volume & issue: Vol. 26, no. 5
p. 106634

Abstract

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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.

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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