tetano
Editor, Senior Moderator
Cell Stem Cell
. 2020 Oct 21;S1934-5909(20)30498-7.
doi: 10.1016/j.stem.2020.10.004. Online ahead of print.
Three-Dimensional Human Alveolar Stem Cell Culture Models Reveal Infection Response to SARS-CoV-2
Jeonghwan Youk[SUP] 1 [/SUP], Taewoo Kim[SUP] 2 [/SUP], Kelly V Evans[SUP] 3 [/SUP], Young-Il Jeong[SUP] 4 [/SUP], Yongsuk Hur[SUP] 5 [/SUP], Seon Pyo Hong[SUP] 6 [/SUP], Je Hyoung Kim[SUP] 4 [/SUP], Kijong Yi[SUP] 2 [/SUP], Su Yeon Kim[SUP] 2 [/SUP], Kwon Joong Na[SUP] 7 [/SUP], Thomas Bleazard[SUP] 8 [/SUP], Ho Min Kim[SUP] 9 [/SUP], Mick Fellows[SUP] 10 [/SUP], Krishnaa T Mahbubani[SUP] 11 [/SUP], Kourosh Saeb-Parsy[SUP] 11 [/SUP], Seon Young Kim[SUP] 12 [/SUP], Young Tae Kim[SUP] 13 [/SUP], Gou Young Koh[SUP] 14 [/SUP], Byeong-Sun Choi[SUP] 15 [/SUP], Young Seok Ju[SUP] 16 [/SUP], Joo-Hyeon Lee[SUP] 17 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is the cause of a present pandemic, infects human lung alveolar type 2 (hAT2) cells. Characterizing pathogenesis is crucial for developing vaccines and therapeutics. However, the lack of models mirroring the cellular physiology and pathology of hAT2 cells limits the study. Here, we develop a feeder-free, long-term, three-dimensional (3D) culture technique for hAT2 cells derived from primary human lung tissue and investigate infection response to SARS-CoV-2. By imaging-based analysis and single-cell transcriptome profiling, we reveal rapid viral replication and the increased expression of interferon-associated genes and proinflammatory genes in infected hAT2 cells, indicating a robust endogenous innate immune response. Further tracing of viral mutations acquired during transmission identifies full infection of individual cells effectively from a single viral entry. Our study provides deep insights into the pathogenesis of SARS-CoV-2 and the application of defined 3D hAT2 cultures as models for respiratory diseases.
Keywords: 3D cultures; COVID-19; SARS-CoV-2; alveolar stem cells; electron microscopy; human alveolar type 2 cells; infection; interferon; interferon-stimulating genes; single-cell RNA-seq
. 2020 Oct 21;S1934-5909(20)30498-7.
doi: 10.1016/j.stem.2020.10.004. Online ahead of print.
Three-Dimensional Human Alveolar Stem Cell Culture Models Reveal Infection Response to SARS-CoV-2
Jeonghwan Youk[SUP] 1 [/SUP], Taewoo Kim[SUP] 2 [/SUP], Kelly V Evans[SUP] 3 [/SUP], Young-Il Jeong[SUP] 4 [/SUP], Yongsuk Hur[SUP] 5 [/SUP], Seon Pyo Hong[SUP] 6 [/SUP], Je Hyoung Kim[SUP] 4 [/SUP], Kijong Yi[SUP] 2 [/SUP], Su Yeon Kim[SUP] 2 [/SUP], Kwon Joong Na[SUP] 7 [/SUP], Thomas Bleazard[SUP] 8 [/SUP], Ho Min Kim[SUP] 9 [/SUP], Mick Fellows[SUP] 10 [/SUP], Krishnaa T Mahbubani[SUP] 11 [/SUP], Kourosh Saeb-Parsy[SUP] 11 [/SUP], Seon Young Kim[SUP] 12 [/SUP], Young Tae Kim[SUP] 13 [/SUP], Gou Young Koh[SUP] 14 [/SUP], Byeong-Sun Choi[SUP] 15 [/SUP], Young Seok Ju[SUP] 16 [/SUP], Joo-Hyeon Lee[SUP] 17 [/SUP]
Affiliations
- PMID: 33142113
- DOI: 10.1016/j.stem.2020.10.004
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is the cause of a present pandemic, infects human lung alveolar type 2 (hAT2) cells. Characterizing pathogenesis is crucial for developing vaccines and therapeutics. However, the lack of models mirroring the cellular physiology and pathology of hAT2 cells limits the study. Here, we develop a feeder-free, long-term, three-dimensional (3D) culture technique for hAT2 cells derived from primary human lung tissue and investigate infection response to SARS-CoV-2. By imaging-based analysis and single-cell transcriptome profiling, we reveal rapid viral replication and the increased expression of interferon-associated genes and proinflammatory genes in infected hAT2 cells, indicating a robust endogenous innate immune response. Further tracing of viral mutations acquired during transmission identifies full infection of individual cells effectively from a single viral entry. Our study provides deep insights into the pathogenesis of SARS-CoV-2 and the application of defined 3D hAT2 cultures as models for respiratory diseases.
Keywords: 3D cultures; COVID-19; SARS-CoV-2; alveolar stem cells; electron microscopy; human alveolar type 2 cells; infection; interferon; interferon-stimulating genes; single-cell RNA-seq