Establishment of Pancreatic Cancer-Derived Tumor Organoids and Fibroblasts From Fresh Tissue
Creators
- Jesús Frutos Díaz-Alejo1
- Simon April-Monn2
- Marina Cihova3
- Verona Buocikova3
- Jorge Villalón López4
- Maria Urbanova3
- Carmen G. Lechuga5
- Miroslav Tomas6
- Peter Dubovan6
- Bárbara Luna Sánchez7
- Sonia Camaño Páez7
- Alfonso Sanjuanbenito8
- Eduardo Lobo9
- Estefanía Romio de la Heras10
- Carmen Guerra11
- Carolina de la Pinta12
- Emma Barreto Melian13
- Mercedes Rodríguez Garrote13
- Alfredo Carrato14
- Laura Ruiz-Cañas15
- Bruno Sainz, Jr.16
- Ana Torres7
- Bozena Smolkova3
- Julie Earl17
- 1. Molecular Epidemiology and Predictive Tumor Markers Group, Area 3, Ramón y Cajal Health Research Institute (IRYCIS); The Biomedical Research Network in Cancer (CIBERONC); Biobank and Biomodels Platform (PT20/0045), ISCIII research and development platforms in biomedicine and health sciences, BioBank Hospital Ramón y Cajal-IRYCIS, Spanish National Biobanks Network (ISCIII Biobank Register No. B.0000678), Ramón y Cajal Health Research Institute (IRYCIS); Faculty of Medicine, University of Alcalá de Henares
- 2. Institute of Tissue Medicine and Pathology, University of Bern
- 3. Department of Molecular Oncology, Cancer Research Institute, Biomedical Research Center of the Slovak Academy of Sciences
- 4. Molecular Epidemiology and Predictive Tumor Markers Group, Area 3, Ramón y Cajal Health Research Institute (IRYCIS); Biobank and Biomodels Platform (PT20/0045), ISCIII research and development platforms in biomedicine and health sciences, BioBank Hospital Ramón y Cajal-IRYCIS, Spanish National Biobanks Network (ISCIII Biobank Register No. B.0000678), Ramón y Cajal Health Research Institute (IRYCIS)
- 5. Experimental Oncology, Molecular Oncology Program, Centro Nacional de Investigaciones Oncológicas (CNIO)
- 6. Department of Molecular Oncology, Cancer Research Institute, Biomedical Research Center of the Slovak Academy of Sciences; Department of Surgical Oncology, National Cancer Institute, Slovak Medical University
- 7. Biobank and Biomodels Platform (PT20/0045), ISCIII research and development platforms in biomedicine and health sciences, BioBank Hospital Ramón y Cajal-IRYCIS, Spanish National Biobanks Network (ISCIII Biobank Register No. B.0000678), Ramón y Cajal Health Research Institute (IRYCIS)
- 8. The Biomedical Research Network in Cancer (CIBERONC); Pancreatic and Biliopancreatic Surgery Unit, Hospital Universitario Ramón y Cajal
- 9. Pancreatic and Biliopancreatic Surgery Unit, Hospital Universitario Ramón y Cajal
- 10. Department of Pathology, Hospital Universitario Ramón y Cajal
- 11. The Biomedical Research Network in Cancer (CIBERONC); Experimental Oncology, Molecular Oncology Program, Centro Nacional de Investigaciones Oncológicas (CNIO)
- 12. Department of Radiation Oncology, Hospital Universitario Ramón y Cajal
- 13. Molecular Epidemiology and Predictive Tumor Markers Group, Area 3, Ramón y Cajal Health Research Institute (IRYCIS); The Biomedical Research Network in Cancer (CIBERONC)
- 14. Molecular Epidemiology and Predictive Tumor Markers Group, Area 3, Ramón y Cajal Health Research Institute (IRYCIS); The Biomedical Research Network in Cancer (CIBERONC); Faculty of Medicine, University of Alcalá de Henares
- 15. Biobank and Biomodels Platform (PT20/0045), ISCIII research and development platforms in biomedicine and health sciences, BioBank Hospital Ramón y Cajal-IRYCIS, Spanish National Biobanks Network (ISCIII Biobank Register No. B.0000678), Ramón y Cajal Health Research Institute (IRYCIS); Department of Cancer, Instituto de Investigaciones Biomédicas "Alberto Sols" (IIBM); Cancer Stem Cell and Fibroinflammatory Group, Chronic Diseases and Cancer, Area 3, IRYCIS
- 16. The Biomedical Research Network in Cancer (CIBERONC); Biobank and Biomodels Platform (PT20/0045), ISCIII research and development platforms in biomedicine and health sciences, BioBank Hospital Ramón y Cajal-IRYCIS, Spanish National Biobanks Network (ISCIII Biobank Register No. B.0000678), Ramón y Cajal Health Research Institute (IRYCIS); Department of Cancer, Instituto de Investigaciones Biomédicas "Alberto Sols" (IIBM); Cancer Stem Cell and Fibroinflammatory Group, Chronic Diseases and Cancer, Area 3, IRYCIS
- 17. Molecular Epidemiology and Predictive Tumor Markers Group, Area 3, Ramón y Cajal Health Research Institute (IRYCIS); The Biomedical Research Network in Cancer (CIBERONC); Biobank and Biomodels Platform (PT20/0045), ISCIII research and development platforms in biomedicine and health sciences, BioBank Hospital Ramón y Cajal-IRYCIS, Spanish National Biobanks Network (ISCIII Biobank Register No. B.0000678), Ramón y Cajal Health Research Institute (IRYCIS)
Description
Tumor organoids are three-dimensional (3D) ex vivo tumor models that recapitulate the biological key features of the original primary tumor tissues. Patient-derived tumor organoids have been used in translational cancer research and can be applied to assess treatment sensitivity and resistance, cell-cell interactions, and tumor cell interactions with the tumor microenvironment. Tumor organoids are complex culture systems that require advanced cell culture techniques and culture media with specific growth factor cocktails and a biological basement membrane that mimics the extracellular environment. The ability to establish primary tumor cultures highly depends on the tissue of origin, the cellularity, and the clinical features of the tumor, such as the tumor grade. Furthermore, tissue sample collection, material quality and quantity, as well as correct biobanking and storage are crucial elements of this procedure. The technical capabilities of the laboratory are also crucial factors to consider. Here, we report a validated SOP/protocol that is technically and economically feasible for the culture of ex vivo tumor organoids from fresh tissue samples of pancreatic adenocarcinoma origin, either from fresh primary resected patient donor tissue or patient-derived xenografts (PDX). The technique described herein can be performed in laboratories with basic tissue culture and mouse facilities and is tailored for wide application in the translational oncology field.
Notes
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