Open AccessA Millifluidic Perfusion Cassette for Studying the Pathogenesis of Enteric Infections Using Ex-Vivo Organoids
Author(s) -
Reid Wilson,
Sarah Hewes,
Anubama Rajan,
Sheng-Fung Lin,
Carolyn Bomidi,
Tsuyoshi Iida,
Mary K. Estes,
Anthony W. Maresso,
K. Jane Grande-Allen
Publication year - 2021
Publication title -
annals of biomedical engineering
Language(s) - English
Resource type - Journals
eISSN - 1573-9686
pISSN - 0090-6964
DOI - 10.1007/s10439-020-02705-8
Subject(s) - ex vivo , organoid , fastidious organism , biology , cell culture , in vivo , microbiology and biotechnology , stem cell , computational biology , bacteria , genetics
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.