Dual and Triple Epithelial Co-culture Model Systems with Donor-Derived Microbiota and THP-1 Macrophages To Mimic Host-Microbe Interactions in the Human Sinonasal Cavities
Scope of the method
- Human health
- Translational - Applied Research
- In vitro - Ex vivo
- Other (e.g. bacteria)
Description
- upper respiratory tract
- host-microbe interaction
- air-liquid interface
- sinonasal cavities
- chronic rhinosinusitis
- epithelial barrier function
- macrophages
- host-microbiome interaction
- biotechnology
- microbiology
- immunomodulation
This is a method to study host-microbe interaction in the upper respiratory tract. A physiologically representative epithelial structure, with mucin producing and ciliated cells, is obtained by culturing respiratory epithelial cells at air-liquid interface in Transwell inserts. Optionally, macrophage-like cells, derived from monocytes, can be included to examine immunomodulation. This co-culture system can be apically inoculated with pure strains, a defined mixture of bacteria, or donor-derived nasal microbiota. During host-microbe co-culture, typically 72 h, bacterial adhesion, growth and community composition can be measured, as well as host responses such as cytokine release and epithelial barrier functionality.
- Biosafety cabinet ;
- Incubator ;
- Flow cytometer ;
- Plate-reader ;
- Electrode to measure transepithelial electrical resistance ;
- Micropipettes.
- Still in development
- History of use
- Internally validated
- Published in peer reviewed journal
Pros, cons & Future potential
- - Low-tech ;
- - High throughput ;
- - Commercially available culture system (Transwell) ;
- - Easy sampling ;
- - Variety of samples ;
- - Versatility of host and microbial materials that can be used ;
- - Robust co-culture preserving viability of host cells and bacteria over multiple days.
- - Labour intensive ;
- - Static co-culture (accumulation of metabolites, medium acidification) ;
- - Several weeks required for differentiation ;
- - Low biomass samples of microbial community.
- - Inclusion of more/other host cell types ;
- - Downscaling ;
- - Increasing throughput ;
- - Standardized inoculum.
- - Testing of environmental contaminants (cigarette smoke) ;
- - Antibiotics ;
- - Live biotherapeutic products ;
- - Topical treatments for upper respiratory tract diseases
References, associated documents and other information
De Rudder C, Calatayud Arroyo M, Lebeer S, Van de Wiele T. 2020. Dual and triple epithelial coculture model systems with donorderived microbiota and THP-1 macrophages to mimic host-microbe interactions in the human sinonasal cavities. mSphere 5:e00916-19. https://doi.org/10.1128/mSphere.00916-19.
Corresponding author: Tom.VandeWiele@UGent.be
Contact person
Charlotte De RudderOrganisations
Ghent University (UGent)BW25 - Biotechnology
Belgium
