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P2-218 Human Gut-on-a-Chip as a Model for Enteric Virus Culture

Monday, July 27, 2015
Exhibit Hall (Oregon Convention Center)
Samantha Q. Wales , U.S. Food and Drug Administration-CFSAN , Laurel , MD
Remi Villenave , Wyss Institute for Biologically Inspired Engineering , Boston , MA
Tiama Hamkins-Indik , Wyss Institute for Biologically Inspired Engineering , Boston , MA
Riccardo Barrile , Wyss Institute for Biologically Inspired Engineering , Boston , MA
Efstathia Papafragkou , U.S. Food and Drug Administration-CFSAN , Laurel , MD
Anthony Bahinski , Wyss Institute for Biologically Inspired Engineering , Boston , MA
Michael Kulka , U.S. Food and Drug Administration , Laurel , MD
Donald E. Ingber , Wyss Institute for Biologically Inspired Engineering at Harvard , Boston , MA
Introduction:  The successful cultivation of foodborne viruses is a high-priority research outcome with respect to food safety.   A reproducible cell culture model for human noroviruses (NoV) has been unattainable despite numerous attempts in many cell lines.  The human gut-on-a-chip technology developed by the Wyss Institute offers a highly differentiated model that reconstitutes many features of normal small intestine physiology; namely, villi formation, establishment of tight junctions, and formation of basal proliferative crypts, and therefore provides a good platform in which to study enteric virus replication. 

Purpose: To date, the gut-on-a-chip has never been tested for virus infection.  The objective of these studies was to establish whether the gut-on-a-chip environment could support enteric virus replication and be used as a model for human norovirus culture.

Methods:  The enteric picornavirus Coxsackievirus B1 (CVB1) was tested as a positive control in the system.  CVB1 was adsorbed on the apical or basal side of 5-day differentiated Caco-2 cells in the gut-on-a-chip for 2 h without flow, at which time the cells were rinsed with medium, and continuous flow was resumed.  Apical and basal effluents were collected at 6, 24, and 48 h post-infection (hpi) and assayed for CVB1 production by qRT-PCR and plaque assay.  The cells on the chips were fixed and analyzed by immunofluorescence. 

Results:  CVB1 infection of Caco-2 cells in the gut-on-a-chip resulted in complete destruction of villi and production of infectious virus within 48 hpi, thus demonstrating its utility as a model for enteric virus replication.

Significance:  As the efforts for identifying an appropriate cell culture system that will support NoV replication continue, we have identified and present preliminary data that show the human gut-on-a-chip system is capable of supporting virus replication using CVB1 as a prototype.  Ongoing studies are examining replication of human norovirus in this model.

Back to: Poster Session 2 – Laboratory and Detection Methods; Meat, Poultry and Eggs; Dairy and Beverages; Food Law and Regulation; General Microbiology; Modeling and Risk Assessment
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