Optimized human cell products for research
and drug discovery

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 Specification

Product ID:
Def-INTESTINAL WT
Format:
Cryopreserved
Application:
Research and Drug Discovery

Organoid Morphology

Organoid morphology of Cryo Def-INTESTINAL cells post-thaw and recovery.

Organoid Morphology

Figure 1. Organoids grown encapsulated in matrigel in a 24-well plate.

Immunocytochemistry Analysis

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).

Immunocytochemistry Analysis

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

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

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

CYP3A4 Intestinal Marker

Figure 5. Gene expression analysis shows cytochrome CYP3A4 expression profiles relative to control.

Detection of cytochrome CYP2D6 via qPCR

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

Detection of transporter SLCO2B1 via qPCR

Figure 7. Gene expression analysis shows transporter of SLCO2B1 expression profiles relative to control. 

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