A Microwell-Based Intestinal Organoid-Macrophage Co-Culture System to Study Intestinal Inflammation

Panagiota Kakni, Roman Truckenmüller, Pamela Habibović, Martijn van Griensven, Stefan Giselbrecht*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review


The mammalian intestinal epithelium contains more immune cells than any other tissue, and this is largely because of its constant exposure to pathogens. Macrophages are crucial for maintaining intestinal homeostasis, but they also play a central role in chronic pathologies of the digestive system. We developed a versatile microwell-based intestinal organoid-macrophage co-culture system that enables us to recapitulate features of intestinal inflammation. This microwell-based platform facilitates the controlled positioning of cells in different configurations, continuous in situ monitoring of cell interactions, and high-throughput downstream applications. Using this novel system, we compared the inflammatory response when intestinal organoids were co-cultured with macrophages versus when intestinal organoids were treated with the pro-inflammatory cytokine TNF-α. Furthermore, we demonstrated that the tissue-specific response differs according to the physical distance between the organoids and the macrophages and that the intestinal organoids show an immunomodulatory competence. Our novel microwell-based intestinal organoid model incorporating acellular and cellular components of the immune system can pave the way to unravel unknown mechanisms related to intestinal homeostasis and disorders.

Original languageEnglish
Article number15364
Number of pages17
JournalInternational journal of molecular sciences
Issue number23
Publication statusPublished - 6 Dec 2022


  • Animals
  • Coculture Techniques
  • Organoids
  • Intestines
  • Intestinal Mucosa/pathology
  • Macrophages
  • Mammals


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