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Generation and Analysis of Mouse Intestinal Tumors and Organoids Harboring APC and K-Ras Mutations

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1267)

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.

Key words

Intestine Lgr5 Stem-cells Cancer APC K-Ras Organoids 

Notes

Acknowledgements

The authors like to thank Harry Begthel, Marc van de Wetering, Stieneke van den Brink, and Jeroen Korving for their help in preparing this book chapter.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Hubrecht Institute-KNAW & University Medical Center UtrechtUtrechtThe Netherlands

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