Analysis of Cell Death Induction in Intestinal Organoids In Vitro

  • Thomas Grabinger
  • Eugenia Delgado
  • Thomas Brunner
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1419)

Abstract

The intestinal epithelium has an important function in the absorption of nutrients contained in the food. Furthermore, it also has an important barrier function, preventing luminal pathogens from entering the bloodstream. This single cell layer epithelium is quite sensitive to various cell death-promoting triggers, including drugs, irradiation, and TNF family members, leading to loss of barrier integrity, epithelial erosion, inflammation, malabsorption, and diarrhea. In order to assess the intestinal epithelium-damaging potential of treatments and substances specific test systems are required. As intestinal tumor cell lines are a poor substitute for primary intestinal epithelial cells, and in vivo experiments in mice are costly and often unethical, the use of intestinal organoids cultured from intestinal crypts provide an ideal tool to study cell death induction and mechanisms in primary intestinal epithelial cells. This protocol describes the isolation and culture of intestinal organoids from murine small intestinal crypts, and the quantitative assessment of cell death induction in these organoids.

Key words

Apoptosis Intestinal epithelial cells Crypts TNFα Chemotherapy MTT Irradiation Organoids Enteroids 

Notes

Acknowledgements

This work was supported by Research Grants from the German Science Foundation to TB. Thomas Grabinger received a fellowship from the RTG 1331 graduate school (supported by the German Science Foundation).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Thomas Grabinger
    • 1
  • Eugenia Delgado
    • 1
  • Thomas Brunner
    • 1
  1. 1.Biochemical PharmacologyUniversity of KonstanzKonstanzGermany

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