Differentiating Enteric Glial Cells Within Intestinal Organoids Derived from Human Pluripotent Stem Cells

Area of Science:

• Stem cell biology
• Gastroenterology
• Neuroscience

Background:

• Human intestinal organoids (HIOs) are crucial for studying the gastrointestinal tract.
• Enteric glial cells play a vital role in gut function and development.
• Current methods for co-culturing HIOs and enteric glial cells are limited.

Purpose of the Study:

• To present a novel method for generating and co-culturing human intestinal organoids (HIOs) and enteric glial cells (EGCs) from human pluripotent stem cells.
• To describe techniques for collecting and staining EGCs within HIOs for detailed analysis.
• To provide a reproducible protocol for advancing the study of human EGCs in a relevant in vitro model.

Main Methods:

• Generation of HIOs from human pluripotent stem cells.
• Isolation and differentiation of vagal neural crest cells for EGC generation.
• Co-culture of HIOs and EGCs within Matrigel.
• Immunohistochemical staining of EGCs within the organoid structure.

Main Results:

• Successful generation of both HIOs and EGCs from human pluripotent stem cells.
• Establishment of a co-culture system allowing for the study of HIO-EGC interactions.
• Demonstration of effective methods for visualizing and analyzing EGCs within the organoid model.

Conclusions:

• The presented method enables the generation and co-culture of HIOs and EGCs, providing a valuable tool for research.
• This protocol facilitates the study of human enteric glial cells in a physiologically relevant context.
• The findings offer new avenues for investigating gastrointestinal development and diseases involving the enteric nervous system.