建立和表征南非患者来源的肠道类器官的方案。

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

作者信息

Abdulla Naaziyah, Aronson Ruth, Plessis Thea-Leonie du, Bebington Brendan, Kaur Mandeep

机构信息

School of Molecular and Cell Biology, Faculty of Science, University of the Witwatersrand, Private Bag 3, Johannesburg, South Africa; Infectious Diseases and Oncology Research Institute (IDORI), Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.

School of Molecular and Cell Biology, Faculty of Science, University of the Witwatersrand, Private Bag 3, Johannesburg, South Africa.

出版信息

STAR Protoc. 2025 Jul 22;6(3):103970. doi: 10.1016/j.xpro.2025.103970.

DOI:10.1016/j.xpro.2025.103970

PMID:40705597

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12305567/

Abstract

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.

摘要

患者来源的类器官对于模拟疾病发病机制、进行治疗筛选以及推进个性化医疗具有重要价值。在此，我们展示了一种从南非患者的健康和癌组织中生成并表征肠道类器官的方案。我们描述了生成类器官培养物、定量逆转录聚合酶链反应（RT-qPCR）、免疫荧光显微镜检查和组织学染色的步骤。值得注意的是，我们提供了一种在类器官和组织中进行胆固醇谱分析的详细方法，为在类器官系统中探索脂质生物学的研究人员提供了一个强大的工具。