Protocol for the establishment and characterization of South African patient-derived intestinal organoids.

Patient-derived organoids are valuable for modeling disease pathogenesis, therapeutic screening, and advancing personalized medicine. Here, we present a protocol for generating and characterizing intestinal organoids from South African patients, using both healthy and cancerous tissues. We describe steps for generating organoid cultures, quantitative reverse-transcription polymerase chain reaction (RT-qPCR), immunofluorescence microscopy, and histological staining. Notably, we provide a detailed method for cholesterol profiling in organoids and tissues, offering a powerful tool for researchers exploring lipid biology in organoid systems.