Project
393-Fujimoto-CoV2Infect
Title
SARS-CoV-2 infection to the co-culture of bronchial organoids and vascular beds in a microfluidic device
Description
Blood vessels show various COVID-19-related conditions including thrombosis and cytokine propagation. Existing in vitro blood vessel models cannot represent the consequent changes in the vascular structure or determine the initial infection site, making it difficult to evaluate how epithelial and endothelial tissues are damaged. Here, the authors developed a microphysiological system (MPS) that co-culture the bronchial organoids and the vascular bed to analyze infection site and interactions. In this system, virus-infected organoids caused damage in vascular structure. However, vasculature was not damaged or infected when the virus was directly introduced to vascular bed. The knockout of interferon-related genes and inhibition of the JAK/STAT pathway reduced the vascular damage, indicating the protective effect of interferon response suppression. The results demonstrate selective infection of bronchial epithelial cells and vascular damage by cytokines and also indicate the applicability of MPS to investigate how the infection influences vascular structure and functions.
Funding
KAKENHI (Grant-in-Aid for Scientific Research) on Priority Areas ‘Systems Genomics’ [17017038]
Title
Z-series images of actin filaments in WT-VB in the SARS-CoV-2-infected device culturing KO-BO
Description
Z-series images of vascular bed in the SARS-CoV-2-infected device in which vascular bed and KO-BO were co-culured. KO-BO is bronchial organoids in which Interferon beta (IFNB1) was knocked out.
Channel1: DAPI, Channel2: Alexa Fluor 647 labeled phalloidin, Channel3: GFP, Channel4: ICAM1, Channel5: Differential interference image
Description
Co-culture of bronchial organoids, which were derived from NHBEs (normal human bronchial epithelial
cells), and vascular beds, which were composed from GFP-Expressing Human Umbilical Vein Endothelial Cells, in a special microfluidic device. Interferon beta (IFNB1) was knocked out in the NHBEs.
Intrinsic variables
Interferon beta (IFNB1) was knocked out in NHBEs which is the component of bronchial organoids.
Extrinsic variables
The SARS-CoV-2 virus was applied to the bronchial organoids
Gene ontology: Biological processes
Gene ontology: Cellular components
Gene ontology: Molecular functions
Protein names
Intercellular adhesion molecule 1 (
P05362)
Actin, cytoplasmic 1 (
P60709)
Dimensions
248x250x30x5x1
Visualization methods
primary antibody plus labeled secondary antibody
GFP,Fig4F
4',6-diamidino-2-phenylindole (DAPI)
Contrast enhancing methods
Resolution enhancing methods
Sample preparation methods
fixation method
Body
Olympus-Evident FV3000
Objective
Olympus-Evident UPLXAPO10X