The Intestinal Epithelium Organoid Cultures
- Challenges in establishing long-term systems for intestinal epithelium cultures
- Establishment of long-term gastrointestinal epithelium organoid culture
- The use of small molecules in intestinal organoid culture
- Building of gut organoids
- Cytokines used in long-term gastrointestinal epithelium culture
- Small molecules and peptides used in gastrointestinal culture
Challenges in establishing long-term systems for intestinal epithelium cultures
Establishment of long-term gastrointestinal epithelium organoid culture
1. Intestinal epithelium organoid cultures
- The ability to source normal epithelial cell lines from the stem cells found in the base of intestinal crypts has allowed for the establishment of long-term systems for intestinal epithelium cultures.
- This advancement has allowed for the exploitation of stem cells in tissue regenerative therapies, and the development of treatment models targeting degenerative disorders of the digestive tract.
- The model for a robust, long-term small intestinal epithelium organoid culture system was developed in 2009 .
- Lgr5+ stem cells isolated from murine crypts were cultured with ROCK inhibitor (Y-27632) and the ENR growth factor bundle of EGF (Epidermal Growth Factor), Noggin, and R-Spondin-1.
- This culture system mimics normal intestinal epithelial growth and differentiation and can maintain these characteristics for more than eight months.
2. Human small intestinal epithelium and human colonic epithelial organoid cultures
- Subsequently, a protocol for long-term organoid culturing of the human small intestinal epithelium, and both murine and human colonic epithelia was established.
- This system added the necessary signalling protein Wnt-3a to the above-mentioned ENR growth factor bundle (WENR).
3. Human small intestinal and colonic crypt organoid cultures
- In the case of human small intestinal and colonic crypt cultures, the further addition of p38 MAPK inhibitor (SB202190) and TGF-β inhibitor (A 83-01) was required .
- Currently, these protocols are routinely used in studies involving human or mouse intestinal crypt cultures [3, 4].
4. Long-term gastric gland and human small intestinal epithelium organoid cultures
- The addition of FGF-10 (Fibroblast Growth Factor-10) to the WENR bundle (WENRF) allowed researchers to establish long-term gastric gland and human small intestinal epithelium cultures [5,6,7,8].
The use of small molecules in intestinal organoid cultures
Building of gut organoids
- These gut organoids expanded from healthy, human small intestinal crypts on extracellular matrix, with the assistance of the ENR growth factor bundle, plus Wnt-3a, Y-27632, Gastrin, Nicotinamide, A 83-01, SB 202190 and LY2157299 .
Cytokines used in long-term gastrointestinal epithelium culture
Small molecules and peptides used in gastrointestinal culture
[Poster] Reagents for Organoid Research
[Brochure] Facilitate your research on Gastrointestinal Epithelium Cells with Organoid Cultures
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1. Sato, T, et al. (2009) “Single Lgr5 stem cells build crypt villus structures in vitro without a mesenchymal niche.” Nature 459.7244: 262-265. 2. Sato, T, et al. (2011) “Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium.” Gastroenterology 141.5: 1762-1772. 3. Schwank, G, et al. (2013) “Generation of BAC transgenic epithelial organoids.” PloS One 8.10: e76871. 4. Grabinger, T, et al. (2014) “Ex vivo culture of intestinal crypt organoids as a model system for assessing cell death induction in intestinal epithelial cells and enteropathy.” Cell death & disease 5.5: e1228. 5. Lahar, N, et al. (2011) “Intestinal subepithelial myofibroblasts support in vitro and in vivo growth of human small intestinal epithelium.”PLoS One 6.11: e26898. 6. Jabaji, Z, et al. (2014) “Type I collagen as an extracellular matrix for the in vitro growth of human small intestinal epithelium.” PloS One 9.9: e107814. 7. Barker, N, et al. (2010) “Lgr5+ ve stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro.” Cell Stem Cell 6.1: 25-36. 8. Stange, D, et al. (2013) “Differentiated Troy+ chief cells act as reserve stem cells to generate all lineages of the stomach epithelium.” Cell 155.2: 357-368. 9. Koo, B-K, and Clevers, H, (2014) “Stem cells marked by the R-spondin receptor LGR5.” Gastroenterology 147.2: 289-302. 10. Cramer, J, et al. (2015) “Distinct human stem cell populations in small and large intestine.” PloS One 10.3: e0118792. 11. Farin, H. F., Van Es, J. H., & Clevers, H. (2012) “Redundant sources of Wnt regulate intestinal stem cells and promote formation of Paneth cells.” Gastroenterology 143.6: 1518-1529. 12. Schweinlin, M, et al. (2016). “Development of an advanced primary human in vitro model of the small intestine.” Tissue Engineering Part C: Methods
13.Peprotech. (n.d.). Facilitate your research on Gastrointestinal Epithelium Cells with Organoid Cultures. Retrieved November 16, 2021, from https://www.peprotech.com/download/2024.
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