Bioengineered in vitro enteric nervous system.

Area of Science:

• Neuroscience
• Gastroenterology
• Tissue Engineering

Background:

• Bidirectional communication between the central nervous system and the gastrointestinal tract via the enteric nervous system is crucial for human health but poorly understood.
• Existing in vivo animal models have limitations for studying these complex gut-brain interactions.
• There is a need for advanced 3D human in vitro intestinal models to replicate the intricate cell interactions of the enteric nervous system.

Purpose of the Study:

• To develop and characterize a novel 3D-innervated human intestinal tissue model.
• To investigate the survival, function, and cellular interactions within this in vitro model.
• To establish a tool for studying neural circuits controlling the human intestine.

Main Methods:

• Constructed a 3D intestinal tissue model using human-induced neural stem cells, enterocyte-like (Caco-2) and goblet-like (HT29-MTX) cells, and intestinal myofibroblasts.
• Differentiated neural stem cells into enteric nervous system neural cell types.
• Cultured the model for 5 weeks, assessing cell survival, migration, and gene expression.

Main Results:

• The 3D model supported the survival and function of enteric nervous system cells for over 5 weeks, with evident mucosal and neural transcription factors.
• Human-induced neural stem cells migrated towards luminal epithelial cells, stimulated by neural growth factor.
• The model expressed nNOS-expressing neurons and the TAC1 gene, recapitulating key enteric nervous system phenotypes.

Conclusions:

• The developed 3D-innervated intestinal tissue model successfully recapitulates human enteric nervous system phenotypes.
• This model provides a valuable new tool for understanding neural circuits and communication networks in the human gut.
• It offers a promising platform for studying conditions involving gut-brain axis dysregulation.