A surgical orthotopic organoid transplantation approach in mice to visualize and study colorectal cancer progression

ABSTRACT Most currently available colorectal cancer (CRC) mouse models are not suitable for studying progression toward the metastatic stage. Recently, establishment of tumor organoid lines,

either from murine CRC models or patients, and the possibility of engineering them with genome-editing technologies, have provided a large collection of tumor material faithfully

recapitulating phenotypic and genetic heterogeneity of native tumors. To study tumor progression in the natural _in vivo_ environment, we developed an orthotopic approach based on

transplantation of CRC organoids into the cecal epithelium. The 20-min procedure is described in detail here and enables growth of transplanted organoids into a single tumor mass within the

intestinal tract. Due to long latency, tumor cells are capable of spreading through the blood circulation and forming metastases at distant sites. This method is designed to generate tumors

suitable for studying CRC progression, thereby providing the opportunity to visualize tumor cell dynamics _in vivo_ in real time by intravital microscopy. Access through your institution Buy

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COLON CANCER FOR THE ANALYSIS OF SPORADIC AND INFLAMMATION-DRIVEN TUMOR PROGRESSION AND LYMPH NODE METASTASIS Article 14 December 2020 GASTROINTESTINAL CANCER ORGANOIDS—APPLICATIONS IN BASIC

AND TRANSLATIONAL CANCER RESEARCH Article Open access 18 October 2021 _SLEEPING BEAUTY_ TRANSPOSON MUTAGENESIS IN MOUSE INTESTINAL ORGANOIDS IDENTIFIES GENES INVOLVED IN TUMOR PROGRESSION

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thank A. de Graaff and the Hubrecht Imaging Centre for imaging support. We thank O. Sansom and E. Hong Tan (Beatson Institute, Glasgow, UK) for providing the murine tumor organoid line. This

work was financially supported by a Dutch Cancer Society Fellowship (BUIT-2013-5847 to S.J.E.S.), by the Dutch Cancer Society (KWF)/Alpe d′HuZes Bas Mulder Award (KWF/Alpe d′HuZes 10218, to

J.D.), by European Research Council Grant CANCER-RECURRENCE 648804 (to J.v.R.), by the CancerGenomics.nl (Netherlands Organisation for Scientific Research) program (to J.v.R.), by the

Doctor Josef Steiner Foundation (to J.v.R) and by the European Union′s Horizon 2020 research and innovation program under Marie Sklodowska-Curie grant agreement no. 642866 (to J.v.R). AUTHOR

INFORMATION AUTHORS AND AFFILIATIONS * Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and UMC Utrecht, Utrecht, The Netherlands Arianna Fumagalli, Saskia J E

Suijkerbuijk, Harry Begthel, Evelyne Beerling & Jacco van Rheenen * Molecular Pathology, Oncode Institute, The Netherlands Cancer Institute, Amsterdam, The Netherlands, Utrecht, The

Netherlands Arianna Fumagalli, Saskia J E Suijkerbuijk, Evelyne Beerling & Jacco van Rheenen * Molecular Cancer Research, Center for Molecular Medicine, University Medical Center

Utrecht, Utrecht, The Netherlands Koen C Oost & Hugo J Snippert * Princess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands Jarno Drost Authors * Arianna Fumagalli View

author publications You can also search for this author inPubMed Google Scholar * Saskia J E Suijkerbuijk View author publications You can also search for this author inPubMed Google Scholar

* Harry Begthel View author publications You can also search for this author inPubMed Google Scholar * Evelyne Beerling View author publications You can also search for this author inPubMed

 Google Scholar * Koen C Oost View author publications You can also search for this author inPubMed Google Scholar * Hugo J Snippert View author publications You can also search for this

author inPubMed Google Scholar * Jacco van Rheenen View author publications You can also search for this author inPubMed Google Scholar * Jarno Drost View author publications You can also

search for this author inPubMed Google Scholar CONTRIBUTIONS A.F. developed the orthotopic transplantation technique; A.F. and J.D. performed the experiments; H.B., E.B. and S.J.E.S. helped

with data analysis and preparation of the figures; S.J.E.S edited the video; K.C.O. and H.J.S. provided the patient-derived CRC organoids; J.D. and J.v.R. supervised the study; A.F., J.D.

and J.v.R. wrote the manuscript. CORRESPONDING AUTHORS Correspondence to Jacco van Rheenen or Jarno Drost. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial

interests. SUPPLEMENTARY INFORMATION 41596_2018_BFNPROT2017137_MOESM136_ESM.MP4 Set up and demonstration of the surgical procedure to transplant CRC organoids into the caecal wall of

recipient mice. (MP4 19876 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Fumagalli, A., Suijkerbuijk, S., Begthel, H. _et al._ A surgical

orthotopic organoid transplantation approach in mice to visualize and study colorectal cancer progression. _Nat Protoc_ 13, 235–247 (2018). https://doi.org/10.1038/nprot.2017.137 Download

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