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Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the etiologic agent for the pneumonia outbreak that started in early December 2019 in Wuhan City, Hubei Province, China. To date, coronavirus disease (COVID-19) has caused almost 6 million deaths worldwide. The abili...

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Autores principales: Rosli, Siti Nur Zawani, Dimeng, Sitti Rahmawati, Shamsuddin, Farah, Mohd Ali, Mohammad Ridhuan, Muhamad Hendri, Nur Afrina, Suppiah, Jeyanthi, Mohd Zain, Rozainanee, Thayan, Ravindran, Ahmad, Norazah
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10295990/
https://www.ncbi.nlm.nih.gov/pubmed/37371753
http://dx.doi.org/10.3390/biomedicines11061658
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author Rosli, Siti Nur Zawani
Dimeng, Sitti Rahmawati
Shamsuddin, Farah
Mohd Ali, Mohammad Ridhuan
Muhamad Hendri, Nur Afrina
Suppiah, Jeyanthi
Mohd Zain, Rozainanee
Thayan, Ravindran
Ahmad, Norazah
author_facet Rosli, Siti Nur Zawani
Dimeng, Sitti Rahmawati
Shamsuddin, Farah
Mohd Ali, Mohammad Ridhuan
Muhamad Hendri, Nur Afrina
Suppiah, Jeyanthi
Mohd Zain, Rozainanee
Thayan, Ravindran
Ahmad, Norazah
author_sort Rosli, Siti Nur Zawani
collection PubMed
description Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the etiologic agent for the pneumonia outbreak that started in early December 2019 in Wuhan City, Hubei Province, China. To date, coronavirus disease (COVID-19) has caused almost 6 million deaths worldwide. The ability to propagate the virus into a customizable volume will enable better research on COVID-19 therapy, vaccine development, and many others. In the search for the most efficient replication host, we inoculated three (3) local SARS-CoV-2 isolates of different lineages (Clade L/Lineage B Wuhan, Clade GR/Lineage B.1.1.354, and Clade O/Lineage B.6.2) into various clinically important mammalian cell lines. The replication profile of these isolates was evaluated based on the formation of cytopathic effects (CPE), viral load (Ct value and plaque-forming unit (pfu)), as well as observation by electron microscopy (EM). Next-generation sequencing (NGS) was performed to examine the genomic stability of the propagated SARS-CoV-2 in these cell lines. Our study found that Vero E6 and Vero CCL-81 cell lines posed similar capacities in propagating the local isolates, with Vero CCL-81 demonstrating exceptional potency in conserving the genomic stability of the Lineage B Wuhan isolate. In addition, our study demonstrated the utility of Calu-3 cells as a replication host for SARS-CoV-2 without causing substantial cellular senescence. In conclusion, this study provides crucial information on the growth profile of Malaysian SARS-CoV-2 in various mammalian cell lines and thus will be a great source of reference for better isolation and propagation of the SARS-CoV-2 virus isolated in Malaysia.
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spelling pubmed-102959902023-06-28 Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia Rosli, Siti Nur Zawani Dimeng, Sitti Rahmawati Shamsuddin, Farah Mohd Ali, Mohammad Ridhuan Muhamad Hendri, Nur Afrina Suppiah, Jeyanthi Mohd Zain, Rozainanee Thayan, Ravindran Ahmad, Norazah Biomedicines Article Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the etiologic agent for the pneumonia outbreak that started in early December 2019 in Wuhan City, Hubei Province, China. To date, coronavirus disease (COVID-19) has caused almost 6 million deaths worldwide. The ability to propagate the virus into a customizable volume will enable better research on COVID-19 therapy, vaccine development, and many others. In the search for the most efficient replication host, we inoculated three (3) local SARS-CoV-2 isolates of different lineages (Clade L/Lineage B Wuhan, Clade GR/Lineage B.1.1.354, and Clade O/Lineage B.6.2) into various clinically important mammalian cell lines. The replication profile of these isolates was evaluated based on the formation of cytopathic effects (CPE), viral load (Ct value and plaque-forming unit (pfu)), as well as observation by electron microscopy (EM). Next-generation sequencing (NGS) was performed to examine the genomic stability of the propagated SARS-CoV-2 in these cell lines. Our study found that Vero E6 and Vero CCL-81 cell lines posed similar capacities in propagating the local isolates, with Vero CCL-81 demonstrating exceptional potency in conserving the genomic stability of the Lineage B Wuhan isolate. In addition, our study demonstrated the utility of Calu-3 cells as a replication host for SARS-CoV-2 without causing substantial cellular senescence. In conclusion, this study provides crucial information on the growth profile of Malaysian SARS-CoV-2 in various mammalian cell lines and thus will be a great source of reference for better isolation and propagation of the SARS-CoV-2 virus isolated in Malaysia. MDPI 2023-06-07 /pmc/articles/PMC10295990/ /pubmed/37371753 http://dx.doi.org/10.3390/biomedicines11061658 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rosli, Siti Nur Zawani
Dimeng, Sitti Rahmawati
Shamsuddin, Farah
Mohd Ali, Mohammad Ridhuan
Muhamad Hendri, Nur Afrina
Suppiah, Jeyanthi
Mohd Zain, Rozainanee
Thayan, Ravindran
Ahmad, Norazah
Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia
title Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia
title_full Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia
title_fullStr Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia
title_full_unstemmed Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia
title_short Vero CCL-81 and Calu-3 Cell Lines as Alternative Hosts for Isolation and Propagation of SARS-CoV-2 Isolated in Malaysia
title_sort vero ccl-81 and calu-3 cell lines as alternative hosts for isolation and propagation of sars-cov-2 isolated in malaysia
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10295990/
https://www.ncbi.nlm.nih.gov/pubmed/37371753
http://dx.doi.org/10.3390/biomedicines11061658
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