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

Arianna Fumagalli, Saskia J E Suijkerbuijk, Harry Begthel, Evelyne Beerling, Koen C Oost, Hugo J Snippert, Jacco van Rheenen, Jarno Drost

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Pages (from-to)235-247
Number of pages13
JournalNature protocols
Issue number2
Publication statusPublished - Feb 2018


  • Animals
  • CRISPR-Cas Systems/genetics
  • Cell Transformation, Neoplastic
  • Colonic Neoplasms
  • Colonoscopy/methods
  • Colorectal Neoplasms/physiopathology
  • Disease Models, Animal
  • Gene Editing/methods
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Liver Neoplasms
  • Mice
  • Mice, Inbred NOD
  • Organoids/physiology
  • Xenograft Model Antitumor Assays


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