Intestinal cell morphology
Our intestinal cells grow encapsulated in matrigel and develop crypt-like structures as they mature. Our organoids can typically be passaged 10 times~ weekly passage so have several months usage.
Figure 1. Typical intestinal morphology is observed in Def-INTESTINAL cells. Organoids grown encapsulated in matrigel in a 24 well plate.
Figure 2. Organoids display positive staining of key intestinal cell markers including: epithelial cells (E-cadherin), enterocytes (villin), goblet cells (mucin), enteroendocrine (somatostatin), and Paneth cells (lysozyme).
Key intestinal cell marker analysis
Figure 3. Def-INTESTINAL organoids have been demonstrated to display many key gut markers. Gene expression analysis shows key intestinal markers KRT19, Villin, and CHGA expression profiles relative against primary human control relative to GAPDH.
Figure 4. A range of cytochrome P450 and UGT enzymes have been identified in Def-INTESTINAL organoids. Gene expression analysis shows (A) cytochromes CYP3A4, CYP2D6, CYP2J2and (B) UGT enzyme UGT2B7 expression against primary human control.
Drug transporter analysis
Down-regulation or inhibition of ABC efflux transporters in the intestine can be used as a strategy to improve oral drug bioavailability of known substrates as these transporters prevent drug molecules from being absorbed.
The SLC (solute carrier) family have an important role in physiological processes ranging from the cellular uptake of nutrients to the absorption of drugs and other xenobiotics. SLCs are primarily involved in the uptake of small molecules into cells.
Figure 5. (A) Gene expression analysis shows transporter ABCB1 expression profiles against primary control. (B) Gene expression analysis shows transporter SLCO2B1 expression profiles against primary control.
Figure 6. (A) Def-INTESTINAL organoids can transport Rhodamine 123, a specific substrate of MDR1. MDR1 activity is inhibited by Verapamil a specific inhibitor. (B) Immunostaining of intestinal organoids showing localization of the MDR1 transporter protein within highly folded crypt structures.
Brochure: DefiniGEN Intestinal Organoids
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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