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Protocol for the generation, characterization, and functional assays of organoid cultures from normal and cancer-prone human esophageal tissues

Year: 2024

Type: article

Abstract: Multiple sampling strategies to cover different or dynamic stages of malignant continuum with organoid cultures provide a valuable platform for epithelium homeostasis, transformation, and cancer progression. Here, we present a protocol to initiate, culture, passage, and characterize organoids from normal and cancer-prone human esophageal tissues. We describe steps for multiple sampling of malignant continuum and the initiation and maintenance of multi-stage organoids. We then detail procedures for the histological characterization of organoids and co-culture systems based on organoids and stromal cells. For complete details on the use and execution of this protocol, please refer to Chen et al. 1 • Multiple sampling strategies to cover dynamic stages of malignant continuum • Multi-stage esophageal organoid cultures • Characterizing multi-stage organoids using multiplex immunofluorescence • Co-culture assays of organoids and fibroblasts Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Multiple sampling strategies to cover different or dynamic stages of malignant continuum with organoid cultures provide a valuable platform for epithelium homeostasis, transformation, and cancer progression. Here, we present a protocol to initiate, culture, passage, and characterize organoids from normal and cancer-prone human esophageal tissues. We describe steps for multiple sampling of malignant continuum and the initiation and maintenance of multi-stage organoids. We then detail procedures for the histological characterization of organoids and co-culture systems based on organoids and stromal cells.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103316

Authors: Shaosen Zhang, Guoyu Cheng, Shihao Zhu, Dongxin Lin, Chen Wu