Cell response analysis in SARS-CoV-2 infected bronchial organoids

  1. Emi Sano
  2. Tatsuya Suzuki
  3. Rina Hashimoto
  4. Yumi Itoh
  5. Ayaka Sakamoto
  6. Yusuke Sakai
  7. Akatsuki Saito
  8. Daisuke Okuzaki
  9. Daisuke Motooka
  10. Yukiko Muramoto
  11. Takeshi Noda
  12. Tomohiko Takasaki
  13. Jun-Ichi Sakuragi
  14. Shohei Minami
  15. Takeshi Kobayashi
  16. Takuya Yamamoto
  17. Yasufumi Matsumura
  18. Miki Nagao
  19. Toru Okamoto
  20. Kazuo Takayama

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Abstract

The development of an in vitro cell model that can be used to study severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research is expected. Here we conducted infection experiments in bronchial organoids (BO) and an BO-derived air-liquid interface model (BO-ALI) using 8 SARS-CoV-2 variants. The infection efficiency in BO-ALI was more than 1,000 times higher than that in BO. Among the bronchial epithelial cells, we found that ciliated cells were infected with the virus, but basal cells were not. Ciliated cells died 7 days after the viral infection, but basal cells survived after the viral infection and differentiated into ciliated cells. Fibroblast growth factor 10 signaling was essential for this differentiation. These results indicate that BO and BO-ALI may be used not only to evaluate the cell response to SARS-CoV-2 and coronavirus disease 2019 (COVID-19) therapeutic agents, but also for airway regeneration studies.

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  1. Version published to 10.1038/s42003-022-03499-2
  2. ScreenIT

    SciScore for 10.1101/2020.05.25.115600: (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
    A549 culture: A549 cells were cultured with Ham’s F12 medium (Thermo Fisher Scientific) containing 10% FBS, 1×GlutaMAX (Thermo Fisher Scientific), and penicillin-streptomycin.
    A549
    suggested: None
    The virus was plaque-purified and propagated in Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    SARS-CoV-2 virus plaque assays: VeroE6/TMPRSS2 cells (JCRB1819, JCRB Cell Bank) 17 were seeded on 12 well plates (1.7×105 cells/well) and incubated for 24 hr.
    VeroE6/TMPRSS2
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Software and Algorithms
    SentencesResources
    The trimmed reads were mapped to the human reference genome sequences (hg19) using HISAT2 ver 2.1.0.
    HISAT2
    suggested: (HISAT2, RRID:SCR_015530)
    The raw counts were calculated using featureCounts ver 2.0.0 and used for heatmap visualization with integrated differential expression and pathway analysis (iDEP, (http://ge-lab.org/idep/)) 18.
    featureCounts
    suggested: (featureCounts, RRID:SCR_012919)
    Access to raw data concerning this study was submitted under Gene Expression Omnibus (GEO) accession number GSE150819.
    Gene Expression Omnibus
    suggested: (Gene Expression Omnibus (GEO, RRID:SCR_005012)

    Results from OddPub: Thank you for sharing your data.

    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: No clinical trial numbers were referenced.

    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

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  3. Version published to 10.1101/2020.05.25.115600v2 on bioRxiv
  4. Version published to 10.1101/2020.05.25.115600v1 on bioRxiv