Modeling Epithelial Homeostasis and Perturbation in Three-Dimensional Human Esophageal Organoids

Area of Science:

• Gastroenterology
• Stem Cell Biology
• Organoid Technology

Background:

• Advanced Dulbecco's Modified Eagle Medium-Nutrient Mixture F12 (ADF) is used for esophageal organoids but is suboptimal for normal human esophageal organoids.
• This limitation hinders comparative studies between normal and diseased esophageal organoids.

Purpose of the Study:

• To optimize ADF-based medium for improved generation of normal human esophageal organoids.
• To establish a model for studying human esophageal epithelial pathology and disease.

Main Methods:

• Utilized immortalized esophageal keratinocyte lines (EPC1, EPC2) and esophageal biopsies to generate 3D organoids.
• Optimized medium by assessing epidermal growth factor (EGF) and transforming growth factor-β (TGF-β) signaling.
• Modeled pathology using interleukin-13 (IL-13) or UAB30 to induce epithelial changes.

Main Results:

• Excessive EGF and TGF-β signaling limited normal organoid formation.
• Optimized medium, named HOME0, significantly improved normal human esophageal organoid formation.
• IL-13 and UAB30 in HOME0-grown organoids induced basal cell hyperplasia, mimicking disease states.

Conclusions:

• HOME0 medium facilitates the modeling of human esophageal epithelial homeostasis and disease.
• This optimized system allows for comparative studies of human esophageal organoids with those from other species or organs.
• HOME0 supports the study of esophageal epithelial differentiation gradients and responses to inflammatory or signaling perturbations.