Protocol for culturing olfactory epithelium organoids supporting neuronal differentiation.

Area of Science:

• Neuroscience
• Stem Cell Biology
• Epithelial Biology

Background:

• The olfactory epithelium (OE) harbors stem cells crucial for continuous olfactory sensory neuron regeneration.
• This regenerative capacity makes the OE a significant model for studying adult neurogenesis.
• Understanding OE stem cell dynamics is key to regenerative medicine and neuroscience.

Purpose of the Study:

• To establish a reproducible 3D organoid model derived from olfactory epithelium (OE) stem cells.
• To provide a detailed protocol for generating and culturing these OE organoids.
• To facilitate research into epithelial neurogenesis and olfactory sensory neuron development.

Main Methods:

• Dissection of murine olfactory epithelium (OE).
• Establishment and culturing of 3D OE organoids.
• Immunostaining techniques for organoid analysis.
• Protocol validation in mice without cell sorting.

Main Results:

• A highly robust 3D mouse organoid model from OE cells was successfully developed.
• The protocol details a workflow for OE dissection and organoid culture.
• Guidance on immunostaining OE organoids is provided, demonstrating model utility.

Conclusions:

• The developed 3D OE organoid model offers a powerful tool for studying epithelial neurogenesis.
• This protocol provides a validated method for generating OE organoids, applicable to mice.
• The model system supports research into the mechanisms of olfactory sensory neuron generation and maintenance.