Human Intestinal Cells Wild Type
DefiniGEN Intestinal Gut Organoids
Our Def-Intestinal human cell products are functional intestinal organoids displaying similar morphology to primary intestinal cell 3-D cultures including characteristic crypt structure formation. The Def-Intestinal organoids harbor a mixture of cell types normally present in the primary intestinal epithelial barrier in vivo, including goblet cells, Paneth cells, enterocytes, and enteroendocrine cells. The organoids display a polarized monolayer of epithelial cells, with a clear distinction between the apical and basal sides, and microvilli present on the apical surface. The cells also express a range of key intestinal markers such as mucin 2 (goblet cells), Villin 1 (enterocytes), chromogranin A (enteroendocrine cells), and lysozyme (Paneth cells). Multiple CYP450s and transporters such as PXR and SLCO2B1 are also expressed in the cells. Def-Intestinal products are conveniently provided in a cryopreserved vial format which can be thawed, plated in the clients desired format, and passaged multiple times. Intestinal human cells provide an effective system for studying the intestinal epithelium and its interaction with various stimuli including infectious disease as the intestinal mucosa forms the first line of defence against infections mediated by enteric pathogens such as salmonellae.
- Product ID:
- Def-INTESTINAL WT
- Research and Drug Discovery
Organoid morphology of Cryo Def-INTESTINAL cells post-thaw and recovery.
Figure 1. Organoids grown encapsulated in matrigel in a 24-well plate.
ICC analysis has demonstrated that the Def-INTESTINAL organoids display polarized epithelium and include differentiated cell types with distinct morphologies and specific marker gene expression marker profiles. We have identified cells with robust expression of CHGA, a marker for enteroendocrine cells; MUC2, a marker for goblet cells; and VIL1, which marks the villi of enterocytes. In addition, we observe a subset of organoids containing cells that are enriched for lysozyme staining (Figure 2).
Figure 2. Positive staining of Def-INTESTINAL cells expressing key intestinal cell markers such as lysozyme (LYZ) and Villin with control primary duodenum organoids. Images courtesy of Sasha Mendjan and Alexander Lang, University of Cambridge.
Key Intestinal Cell Marker Analysis
Def-INTESTINAL organoids have been generated which display many key gut markers and can be used to model IBD and infectious disease. The organoids maintained express a range of key gut markers (OLFM4, CHGA, MUC2, Villin, and KRT19). Key intestinal marker expression analysis revealed the specific upregulation of intestinal specific genes such as MUC2.
Detection of key intestinal marker villin via qPCR
Figure 3. Gene expression analysis shows key intestinal marker villin expression profiles relative to control.
Detection of nuclear receptor PXR via qPCR
Figure 4. Gene expression analysis shows nuclear receptor PXR expression profiles relative to control.
Detection of cytochrome CYP3A4 via qPCR
Figure 5. Gene expression analysis shows cytochrome CYP3A4 expression profiles relative to control.
Detection of cytochrome CYP2D6 via qPCR
Figure 6. Gene expression analysis shows cytochrome CYP2D6 expression profiles relative to control.
Detection of transporter SLCO2B1 via qPCR
Figure 7. Gene expression analysis shows transporter of SLCO2B1 expression profiles relative to control.
Interaction of Salmonella enterica Serovar Typhimurium with Intestinal Organoids Derived from Human Induced Pluripotent Stem Cells. Forbester JL, Goulding D, Vallier L, Hannan N, Hale C, Pickard D, Mukhopadhyay S, Dougan G. Infect Immun. 2015 Jul;83(7):2926-34.
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