Dataset Fig5E_VB_baricitinib
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 VB in SARS-CoV-2-infected devices with the administration of baricitinib
Description
Z-series images of actin filaments in vascular bed (VB) in SARS-CoV-2-infected devices with the administration of baricitinib, a JAK/STAT inhibitor. In the 3D culture device, vascular bed and bronchial organoids were co-cultured, and the vitrus was infected to the bronchial organoids. The inhibitor was applied to the VB existing channel.
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 normal human bronchial epithelial
cells, and vascular beds, which were composed from GFP-Expressing Human Umbilical Vein Endothelial Cells, in a special microfluidic device.
Intrinsic variables
The commercial HUVECSs expressing GFP were used.
GFP-HUVECs
(Angio-Proteomie, cAP-0001, Boston, MA, USA)
Extrinsic variables
The SARS-CoV-2 virus was applied to the bronchial organoids,
Baricitinib was applied to the vascular bed channel.
Gene ontology: Biological processes
Gene ontology: Cellular components
Gene ontology: Molecular functions
Protein names
Intercellular adhesion molecule 1 (
P05362)
Actin, cytoplasmic 1 (
P60709)
Dimensions
600x400x20x5x1
Visualization methods
primary antibody plus labeled secondary antibody
GFP
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