Generation, Expansion, and Biobanking of Gastrointestinal Patient-Derived Organoids from Tumor and Normal Tissues

Manuel Cabeza-Segura; Blanca Garcia-Micó; Andrés Cervantes; Josefa Castillo

doi:10.1007/978-1-0716-3730-2_9

Generation, Expansion, and Biobanking of Gastrointestinal Patient-Derived Organoids from Tumor and Normal Tissues

Methods Mol Biol. 2024:2777:123-133. doi: 10.1007/978-1-0716-3730-2_9.

Authors

Manuel Cabeza-Segura^#¹, Blanca Garcia-Micó^#¹, Andrés Cervantes^{2

3}, Josefa Castillo^{4

5

6}

Affiliations

¹ Department of Medical Oncology, Hospital Clínico Universitario de Valencia, INCLIVA, Biomedical Research Institute, University of Valencia, Valencia, Spain.
² Department of Medical Oncology, Hospital Clínico Universitario de Valencia, INCLIVA, Biomedical Research Institute, University of Valencia, Valencia, Spain. andres.cervantes@uv.es.
³ Centro de Investigación Biomédica en Red (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain. andres.cervantes@uv.es.
⁴ Department of Medical Oncology, Hospital Clínico Universitario de Valencia, INCLIVA, Biomedical Research Institute, University of Valencia, Valencia, Spain. pepa.castillo@uv.es.
⁵ Centro de Investigación Biomédica en Red (CIBERONC), Instituto de Salud Carlos III, Madrid, Spain. pepa.castillo@uv.es.
⁶ Department of Biochemistry and Molecular Biology, Universitat de Valencia, Burjassot, Spain. pepa.castillo@uv.es.

^# Contributed equally.

PMID: 38478340
DOI: 10.1007/978-1-0716-3730-2_9

Abstract

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.

Keywords: Adult stem cells; Colorectal cancer; Drug sensitivity assay; Gastric cancer; Normal epithelial tissue; Organoid cryopreservation; Organoid establishment; Organoid expansion; Patient-derived organoids; Tumour tissue.

MeSH terms

Adult
Biological Specimen Banks*
Biopsy
Gastrointestinal Tract
Humans
Neoplasms* / pathology
Organoids