TY - JOUR
T1 - Cell culture systems for isolation of SARS-CoV-2 clinical isolates and generation of recombinant virus
AU - Chen, Da Yuan
AU - Turcinovic, Jacquelyn
AU - Feng, Shuchen
AU - Kenney, Devin J.
AU - Chin, Chue Vin
AU - Choudhary, Manish C.
AU - Conway, Hasahn L.
AU - Semaan, Marc
AU - Close, Brianna J.
AU - Tavares, Alexander H.
AU - Seitz, Scott
AU - Khan, Nazimuddin
AU - Kapell, Sebastian
AU - Crossland, Nicholas A.
AU - Li, Jonathan Z.
AU - Douam, Florian
AU - Baker, Susan C.
AU - Connor, John H.
AU - Saeed, Mohsan
N1 - Publisher Copyright:
© 2023 The Author(s)
PY - 2023/5/19
Y1 - 2023/5/19
N2 - 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.
AB - 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.
KW - Methodology in biological sciences
KW - Virology
UR - https://www.scopus.com/pages/publications/85153284693
UR - https://www.scopus.com/pages/publications/85153284693#tab=citedBy
U2 - 10.1016/j.isci.2023.106634
DO - 10.1016/j.isci.2023.106634
M3 - Article
C2 - 37095858
AN - SCOPUS:85153284693
SN - 2589-0042
VL - 26
JO - iScience
JF - iScience
IS - 5
M1 - 106634
ER -