Esophageal Organoids

The esophagus is a muscular tube that actively passes food from the mouth to the stomach. This organ can be affected by congenital diseases, such as esophageal atresia. In addition, there are several diseases that may afflict people later in life. Some of these include esophageal cancer, gastroesophageal reflux disease (GERD), or a rare disease called achalasia, which affects the muscles of the lower esophagus to prevent the contraction of the organ and the passage of food.

Until recently, research on esophageal development and disease was limited to vertebrate animal models. However, there are some substantial differences between the esophagus of murine and human. The human epithelium is many layers thick, while the mouse esophageal epithelium is only a few layers thick but has a keratinize layer facing the lumen. Furthermore, the human esophagus has submucosal glands that are absent in the murine esophagus. Therefore, the murine esophagus is not an ideal model for disease-related research and for treatment development in particular.

Three-dimensional (3D) culture platforms mimicking in vivo esophageal epithelial tissue architecture ex vivo have emerged as powerful experimental tools for studying esophageal biology in the context of homeostasis and pathology. In the esophagus, the 3D culture system allows to investigate multiple biological processes, including epithelial cell proliferation, differentiation, motility, stress response, and both homotypic and heterotypic cell-cell communications. Cellular interactions involve a variety of cell types (e.g., fibroblasts, endothelial cells, and inflammatory cells) in the esophageal tissue microenvironment under homeostatic and pathologic conditions and are mediated by cell surface molecules as well as extracellular matrix proteins. The ability to experimentally manipulate 3D cultures has greatly enhanced our understanding of the molecular mechanisms and signaling pathways underlying esophageal physiology and pathophysiology.

At OrganoLab, we will provide a relatively simple and well-established step-by-step protocol for direct differentiation of human pluripotent stem cells (hPSCs) into human esophageal organoids and organotypic raft cultures, as well as necessary background on esophagus development and immunofluorescent analysis of the final product to verify the culture identity.

References

1. Whelan K. A. et al.; Esophageal 3D Culture Systems as Modeling Tools in Esophageal Epithelial Pathobiology and Personalized Medicine. Cell Mol Gastroenterol Hepatol. 2018, 5(4): 461-478.
2. Shacham-Silverberg V. et al.; Generation of esophageal organoids and organotypic raft cultures from human pluripotent stem cells. Methods Cell Biol. 2020, 159: 1-22.
3. Trisno, S. L. et al.; Esophageal Organoids from Human Pluripotent Stem Cells Delineate Sox2 Functions during Esophageal Specification. Cell Stem Cell. 2018, 23(4): 501-515.
4. Kasagi, Y. et al.; The Esophageal Organoid System Reveals Functional Interplay Between Notch and Cytokines in Reactive Epithelial Changes. Cell Mol Gastroenterol Hepatol. 2018, 5(3): 333-352.