title: Generation and analysis of mouse intestinal tumors and organoids harboring APC and K-Ras mutations.
abstract: Genetically engineered mouse models of intestinal cancer are experimental systems in which mice are genetically manipulated to develop malignancies in the gastrointestinal tract. These models enable researchers to study the mechanisms of onset, progression, and metastasis of the disease. They also provide a valuable biological system which is suitable for testing (novel) drugs in vivo. Recently, an in vitro culture model has been established in which intestinal epithelial stem cells can grow into three-dimensional, ever-expanding epithelial organoids that retain their original organ identity and genetic stability. This culture system has been applied to diseased epithelia, such as adenoma, adenocarcinoma, and Barrett's epithelium. These organoids can be particularly useful for studying the mechanisms of intestinal tumors and to test (novel) drugs in vitro. Here, we describe our current laboratory protocols to generate and analyze intestinal tumors and organoids harboring APC and K-Ras double mutations.
authors: Hubrecht Institute-KNAW & University Medical Center Utrecht, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands.
authors: Johan H
authors: JH
authors: van Es
authors: Hans
authors: H
authors: Clevers
mesh_terms: D025601:Adenomatous Polyposis Coli Protein / Q000235:genetics*; D000818:Animals; D001947:Breeding; D045744:Cell Line, Tumor; D015925:Cryopreservation; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D060005:Genotyping Techniques; D006801:Humans; D007150:Immunohistochemistry; D019426:Integrases / Q000378:metabolism; D007414:Intestinal Neoplasms / Q000235:genetics* / Q000473:pathology*; D007421:Intestine, Small / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D009154:Mutation; D009940:Organoids / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D013629:Tamoxifen / Q000008:administration & dosage / Q000494:pharmacology; D016707:Tissue Fixation
chemical_list: D025601:Adenomatous Polyposis Coli Protein; D013629:Tamoxifen; D004247:DNA; C045073:Cre recombinase; D019426:Integrases; C495971:Hras protein, mouse; D016283:Proto-Oncogene Proteins p21(ras)