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

Nat Protoc. 2018 Feb;13(2):235-247. doi: 10.1038/nprot.2017.137. Epub 2018 Jan 4.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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
  • Organoids / transplantation*
  • Xenograft Model Antitumor Assays