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

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Cancer Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2777))

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.

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References

  1. Drost J, Clevers H (2018) Organoids in cancer research. Nat Rev Cancer 18(7):407–418

    Article  CAS  PubMed  Google Scholar 

  2. Busslinger GA, Lissendorp F, Franken IA, van Hillegersberg R, Ruurda JP, Clevers H et al (2020) The potential and challenges of patient-derived organoids in guiding the multimodality treatment of upper gastrointestinal malignancies. Open Biol 10(4):190274

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Ooft SN, Weeber F, Dijkstra KK, McLean CM, Kaing S, van Werkhoven E et al (2019) Patient-derived organoids can predict response to chemotherapy in metastatic colorectal cancer patients. Sci Transl Med 11(513):eaay2574

    Article  CAS  PubMed  Google Scholar 

  4. de Witte CJ, Espejo Valle-Inclan J, Hami N, Lõhmussaar K, Kopper O, Vreuls CPH et al (2020) Patient-derived ovarian cancer organoids mimic clinical response and exhibit heterogeneous inter- and intrapatient drug responses. Cell Rep 31(11):107762

    Article  PubMed  Google Scholar 

  5. Bleijs M, van de Wetering M, Clevers H, Drost J (2019) Xenograft and organoid model systems in cancer research. EMBO J 38(15):e101654

    Article  PubMed  PubMed Central  Google Scholar 

  6. Cabeza-Segura M, Garcia-Micò B, La Noce M, Nicoletti GF, Conti V, Filippelli A et al (2023) How organoids can improve personalized treatment in patients with gastro-esophageal tumors. Curr Opin Pharmacol 69:102348

    Article  CAS  PubMed  Google Scholar 

  7. Sato T, Stange DE, Ferrante M, Vries RGJ, van Es JH, van den Brink S et al (2011) Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium. Gastroenterology 141(5):1762–1772

    Article  CAS  PubMed  Google Scholar 

  8. van de Wetering M, Francies HE, Francis JM, Bounova G, Iorio F, Pronk A et al (2015) Prospective derivation of a living organoid biobank of colorectal cancer patients. Cell 161(4):933–945

    Article  PubMed  PubMed Central  Google Scholar 

  9. Bartfeld S, Bayram T, van de Wetering M, Huch M, Begthel H, Kujala P et al (2015) In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection. Gastroenterology 148(1):126–136.e6

    Article  PubMed  Google Scholar 

  10. Seidlitz T, Merker SR, Rothe A, Zakrzewski F, von Neubeck C, Grützmann K et al (2019) Human gastric cancer modelling using organoids. Gut 68(2):207–217

    Article  CAS  PubMed  Google Scholar 

  11. Yan HHN, Siu HC, Law S, Ho SL, Yue SSK, Tsui WY et al (2018) A comprehensive human gastric cancer organoid biobank captures tumor subtype heterogeneity and enables therapeutic screening. Cell Stem Cell 23(6):882–897.e11

    Article  CAS  PubMed  Google Scholar 

  12. Papaccio F, García-Mico B, Gimeno-Valiente F, Cabeza-Segura M, Gambardella V, Gutiérrez-Bravo MF et al (2023) Proteotranscriptomic analysis of advanced colorectal cancer patient derived organoids for drug sensitivity prediction. J Exp Clin Cancer Res 42(1):8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Pleguezuelos-Manzano C, Puschhof J, den Brink S, Geurts V, Beumer J, Clevers H (2020) Establishment and culture of human intestinal organoids derived from adult stem cells. Curr Protoc Immunol 130(1)

    Google Scholar 

  14. Broutier L, Andersson-Rolf A, Hindley CJ, Boj SF, Clevers H, Koo BK et al (2016) Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation. Nat Protoc 11(9):1724–1743

    Article  CAS  PubMed  Google Scholar 

  15. Watanabe K, Ueno M, Kamiya D, Nishiyama A, Matsumura M, Wataya T et al (2007) A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol 25(6):681–686

    Article  CAS  PubMed  Google Scholar 

  16. The Cancer Genome Atlas Network (2012) Comprehensive molecular characterization of human colon and rectal cancer. Nature 487(7407):330–337

    Article  Google Scholar 

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Correspondence to Andrés Cervantes or Josefa Castillo .

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Cabeza-Segura, M., Garcia-Micó, B., Cervantes, A., Castillo, J. (2024). Generation, Expansion, and Biobanking of Gastrointestinal Patient-Derived Organoids from Tumor and Normal Tissues. In: Papaccio, F., Papaccio, G. (eds) Cancer Stem Cells. Methods in Molecular Biology, vol 2777. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3730-2_9

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  • DOI: https://doi.org/10.1007/978-1-0716-3730-2_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3729-6

  • Online ISBN: 978-1-0716-3730-2

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