A robust SARS-CoV-2 replication model in primary human epithelial cells at the air liquid interface to assess antiviral agents

  1. Thuc Nguyen Dan Do
  2. Kim Donckers
  3. Laura Vangeel
  4. Arnab K. Chatterjee
  5. Philippe A. Gallay
  6. Michael D. Bobardt
  7. John P. Bilello
  8. Tomas Cihlar
  9. Steven De Jonghe
  10. Johan Neyts
  11. Dirk Jochmans

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Abstract

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  1. Version published to 10.1016/j.antiviral.2021.105122
  2. Version published to 10.1101/2021.03.25.436907v3 on bioRxiv
  3. ScreenIT

    SciScore for 10.1101/2021.03.25.436907: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Cells and virus isolates: The African monkey kidney cell line VeroE6 tagged green fluorescent protein (VeroE6-GFP, kindly provided by M. van Loock, Janssen Pharmaceutica, Beerse, Belgium) and VeroE6 were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, catalogue no. 41965-039) supplemented with 10% v/v heat-inactivated foetal bovine serum (HI-FBS; HyClone, catalogue no. SV03160.03), 1% v/v sodium bicarbonate 7.5% w/v (NaHCO3; Gibco, catalogue no. 25080-060), and 1% v/v Penicillin-Streptomycin 10000 U/mL (P/S; Gibco, catalogue no. 15140148) at 37°C and 5% CO2.
    VeroE6
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    The generation of virus stocks by serial passaging in Huh-7 and VeroE6 cells were fully reported7,8.
    Huh-7
    suggested: CLS Cat# 300156/p7178_HuH7, RRID:CVCL_0336)
    BavPat1 isolate (passage 2 (P2)) and GHB-03021 isolate (P6 and P7) were used for the air liquid-interface experiment while only the latter was used for standard in vitro assays in VeroE6-GFP cells (P6 and P7) and in Huh7 cells (P9).
    VeroE6-GFP
    suggested: None
    Huh7
    suggested: CLS Cat# 300156/p7178_HuH7, RRID:CVCL_0336)
    Experimental Models: Organisms/Strains
    SentencesResources
    Cells and virus isolates: The African monkey kidney cell line VeroE6 tagged green fluorescent protein (VeroE6-GFP, kindly provided by M. van Loock, Janssen Pharmaceutica, Beerse, Belgium) and VeroE6 were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, catalogue no. 41965-039) supplemented with 10% v/v heat-inactivated foetal bovine serum (HI-FBS; HyClone, catalogue no. SV03160.03), 1% v/v sodium bicarbonate 7.5% w/v (NaHCO3; Gibco, catalogue no. 25080-060), and 1% v/v Penicillin-Streptomycin 10000 U/mL (P/S; Gibco, catalogue no. 15140148) at 37°C and 5% CO2.
    VeroE6-GFP
    suggested: None
    Software and Algorithms
    SentencesResources
    Statistical analysis: All statistical comparisons in the study were performed in GraphPad Prism (GraphPad Software, Inc.).
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04315948Active, not recruitingTrial of Treatments for COVID-19 in Hospitalized Adults
    NCT04385095Active, not recruitingTrial of Inhaled Anti-viral (SNG001) for SARS-CoV-2 (COVID-1…
    NCT04492475CompletedAdaptive COVID-19 Treatment Trial 3 (ACTT-3)

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  4. Version published to 10.1101/2021.03.25.436907v2 on bioRxiv
  5. Version published to 10.1101/2021.03.25.436907v1 on bioRxiv