Publication Date
4-1-2021
Journal
Annals of Biomedical Engineering
DOI
10.1007/s10439-020-02705-8
PMID
33409849
PMCID
PMC8012234
PubMedCentral® Posted Date
4-1-2022
PubMedCentral® Full Text Version
Author MSS
Published Open-Access
yes
Keywords
Adult, Biofilms, Caliciviridae Infections, Cells, Cultured, Escherichia coli, Escherichia coli Infections, Escherichia coli Proteins, Fimbriae Proteins, Gastrointestinal Diseases, Host-Pathogen Interactions, Humans, Intestine, Small, Mucins, Norovirus, Organ Culture Techniques, Organoids, Perfusion, Stem Cells, Virulence Factors, Virus Replication, Escherichia coli, bacteria, norovirus, enteroid, organoid, microfluidic, shear, dynamic, microbiome
Abstract
To generate physiologically-relevant experimental models, the study of enteric diarrheal diseases is turning increasingly to advanced in vitro models that combine ex vivo, stem cell-derived "organoid" cell lines with bioengineered culture environments that expose them to mechanical stimuli, such as fluid flow. However, such approaches require considerable technical expertise with both microfabrication and organoid culture, and are, therefore, inaccessible to many researchers. For this reason, we have developed a perfusion system that is simple to fabricate, operate, and maintain. Its dimensions approximate the volume and cell culture area of traditional 96-well plates and allow the incorporation of fastidious primary, stem cell-derived cell lines with only minimal adaptation of their established culture techniques. We show that infections with enteroaggregative E. coli and norovirus, common causes of infectious diarrhea, in the system display important differences from static models, and in some ways better recreate the pathophysiology of in vivo infections. Furthermore, commensal strains of bacteria can be added alongside the pathogens to simulate the effects of a host microbiome on the infectious process. For these reasons, we believe that this perfusion system is a powerful, yet easily accessible tool for studying host-pathogen interactions in the human intestine.
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