Use of Organoids to Characterize Signaling Pathways in Cancer Initiation

Methods Mol Biol. 2018:1765:315-331. doi: 10.1007/978-1-4939-7765-9_21.

Abstract

The development of intestinal organoid technology has greatly accelerated research in the field of colorectal cancer. Contrary to traditional cancer cell lines, organoids are composed of multiple cell types arranged in 3D structures highly reminiscent of their native tissues. Thus, organoids provide a near-physiological and readily accessible model to study tissue morphogenesis, adult stem cell behavior and tumorigenesis. Here, we provide protocols for establishing intestinal organoid cultures from genetically modified mouse lines and describe methods to overexpress and knockout genes of interest using lentiviral-based approaches.

Keywords: Apc; Intestinal stem cells; Lentiviral infection; Organoid; Wnt; Yap.

MeSH terms

  • Adenomatous Polyposis Coli Protein / genetics
  • Animals
  • Cell Transformation, Neoplastic / pathology*
  • Colon / pathology
  • Colorectal Neoplasms / genetics
  • Colorectal Neoplasms / pathology*
  • Gene Knockout Techniques / instrumentation
  • Gene Knockout Techniques / methods
  • Genetic Vectors / genetics
  • Lentivirus / genetics
  • Mice
  • Mice, Transgenic
  • Neoplasms, Experimental / genetics
  • Neoplasms, Experimental / pathology
  • Organoids / pathology*
  • Rectum / pathology
  • Signal Transduction*
  • Tissue Culture Techniques / instrumentation
  • Tissue Culture Techniques / methods*

Substances

  • Adenomatous Polyposis Coli Protein
  • adenomatous polyposis coli protein, mouse