Department of Medical Oncology, Hospital Clínico Universitario de Valencia, INCLIVA, Biomedical Research Institute, University of Valencia, Valencia, Spain.
Centro de Investigación Biomédica en Red (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain.
Methods Mol Biol. 2024;2777:123-133. doi: 10.1007/978-1-0716-3730-2_9.
Patient-derived organoids (PDOs) generated from adult stem cells present in tissues are invaluable tools for translational cancer research (Drost, Clevers, Nat Rev Cancer 18(7):407-418, 2018). The generation of this 3D cultures is not trivial and requires dedicated procedures. Despite the rapid increase in the use of organoids in cancer research, it is noteworthy that published procedures regarding their generation often lack critical information and standardized protocols remain elusive. Addressing these limitations, the protocol described in this chapter offers an in-depth and comprehensive guide to establishing, expanding, and freezing gastrointestinal PDOs obtained from normal and tumor tissue biopsies. Notably, it also provides valuable insights in the form of tips and tricks to guide and overcome potential challenges that may arise during the procedure.
从组织中提取的成人干细胞衍生的类器官（PDO）是转化癌症研究的宝贵工具（Drost、Clevers，Nat Rev Cancer 18(7):407-418, 2018）。这种 3D 培养物的生成并不简单，需要专门的程序。尽管类器官在癌症研究中的应用迅速增加，但值得注意的是，关于其生成的已发表程序往往缺乏关键信息，标准化协议仍然难以捉摸。为了解决这些限制，本章中描述的方案提供了一个深入和全面的指南，用于建立、扩展和冷冻从正常和肿瘤组织活检中获得的胃肠道 PDO。值得注意的是，它还以技巧和窍门的形式提供了有价值的见解，以指导和克服程序中可能出现的潜在挑战。
