Protocol for developing a mucus-producing gut-on-a-chip model from Caco-2 and HT29-MTX-E12 cells

Area of Science:

• Biomedical Engineering
• Cell Biology
• Gastroenterology

Background:

• Static in vitro gut models lack physiological relevance.
• Organ-on-chip (OoC) technology offers improved simulation of in vivo conditions.
• Replication of 3D structure, mucus production, fluid dynamics, and mechanical forces is crucial for accurate gut modeling.

Purpose of the Study:

• To present a protocol for developing a mucus-producing gut-on-a-chip model.
• To utilize Caco-2 and HT29-MTX-E12 cells for constructing the gut-on-a-chip system.
• To establish methods for assessing the model's viability and barrier integrity.

Main Methods:

• Development of a gut-on-a-chip model using Caco-2 and HT29-MTX-E12 cells.
• Assessment of model viability and barrier integrity.
• Application of various microscopy techniques, including confocal microscopy, for analysis.

Main Results:

• Successful development of a mucus-producing gut-on-a-chip model.
• Demonstrated methods for evaluating the model's physiological relevance and functionality.
• Characterization of the model using advanced imaging techniques.

Conclusions:

• The developed protocol enables the creation of a physiologically relevant gut-on-a-chip model.
• This OoC model closely mimics in vivo gut conditions, including mucus production.
• The protocol provides a valuable tool for studying gut physiology and disease in vitro.