Establishment and Histological Analysis of Esophageal Organoids Modeling the Progression from Normal to Cancerous Tissues

Organoids have emerged as a pivotal tool for advancing the understanding of tumorigenesis and cancer therapy. By generating human organoid models representing different tumor stages and performing histological analyses, it is possible to obtain a deeper understanding of the alterations in cellular morphology, spatial architecture, and the expression of key molecular markers as the tumor progresses. This study presents a comprehensive protocol for the establishment and culture of esophageal squamous cell organoids. Additionally, the protocol outlines methods for assessing the expression patterns and spatial organization of critical molecules within the organoids, utilizing techniques such as fixation, embedding, and staining. Through this protocol, significant changes were identified in the spatial structure of esophageal squamous epithelial cells and in the expression of various tumor biomarkers during tumorigenesis. The protocol facilitates the construction and histological analysis of organoids, enabling researchers to investigate the spatial architecture and molecular alterations of epithelial cells across different stages of tumorigenesis and therapeutic intervention.