A Microwell-Based Intestinal Organoid-Macrophage Co-Culture System to Study Intestinal Inflammation
Abstract
:1. Introduction
2. Results
2.1. Monocultures of Intestinal Organoids and RAW 264.7 Macrophages in Microwell Arrays
2.2. Direct Co-Culture of Intestinal Organoids and RAW 264.7 Macrophages in Comparison with TNF-α Treatment of Intestinal Organoid Monocultures
2.3. TNF-α-Induces Changes in Organoid Morphology
2.4. Indirect Co-Culture of Intestinal Organoids and RAW 264.7 Macrophages
2.5. Luminex Assay for TNF-α-Treated Intestinal Organoids
2.6. Luminex Assay for Direct and Indirect Co-Culture of Intestinal Organoids with Macrophages
3. Discussion
4. Materials and Methods
4.1. Fabrication and Preparation of Microwells for Organoid Culture
4.2. Intestinal Organoid Culture
4.3. Macrophage Culture
4.4. Co-Culture of Intestinal Organoids and Macrophages
4.5. Immunofluorescence and Confocal Microscopy
4.6. Enzyme-Linked Immunosorbent Assay (ELISA)
4.7. Luminex Assay
4.8. Scanning Electron Microscopy (SEM)
4.9. Forskolin Assay
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Kakni, P.; Truckenmüller, R.; Habibović, P.; van Griensven, M.; Giselbrecht, S. A Microwell-Based Intestinal Organoid-Macrophage Co-Culture System to Study Intestinal Inflammation. Int. J. Mol. Sci. 2022, 23, 15364. https://doi.org/10.3390/ijms232315364
Kakni P, Truckenmüller R, Habibović P, van Griensven M, Giselbrecht S. A Microwell-Based Intestinal Organoid-Macrophage Co-Culture System to Study Intestinal Inflammation. International Journal of Molecular Sciences. 2022; 23(23):15364. https://doi.org/10.3390/ijms232315364
Chicago/Turabian StyleKakni, Panagiota, Roman Truckenmüller, Pamela Habibović, Martijn van Griensven, and Stefan Giselbrecht. 2022. "A Microwell-Based Intestinal Organoid-Macrophage Co-Culture System to Study Intestinal Inflammation" International Journal of Molecular Sciences 23, no. 23: 15364. https://doi.org/10.3390/ijms232315364