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Intestinal 3D Organoids

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info@definigen.com
UK +44 (0)1223 497106
USA 617-674-3260

DefiniGEN’s iPSC-derived intestinal organoids provide a unique in vitro system to model the human intestine. The organoids display a polarized epithelium and 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 cells can be used for drug absorption, metabolism, induction of transporters, and the modelling of infectious disease.

  • Highly standardized cell product containing human intestinal organoids with consistent performance and biologically relevant data
  • Wild-type donor genetics and karyotype verified
  • Multiple cell types identified in the intestinal organoids including epithelial cells, enterocytes, goblet cells, enteroendocrine cells, and Paneth cells
  • Cells display multiple key gut markers OLFM4, CHGA, MUC2, Villin and KRT19
  • Cells display CYP450 induced activities
  • Organoids can be maintained long-term in culture through passaging

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.

iPSC-derived intestinal organoids display typical crypt-like structures and morphology in culture

Figure 1. Typical intestinal morphology is observed in Def-INTESTINAL cells. Organoids grown encapsulated in matrigel in a 24 well plate.

Immunocytochemistry analysis

iPSC-derived organoids exhibit expression of key intestinal markers; E-cadherin, villin, mucin, somatostatin and lysozyme

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

iPSC-derived organoids express key gut markers such as KRT19, villin and CHGA

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.

(A)

A range of cytochrome P450 enzymes have been identified in Def-INTESTINAL organoids

(B)

A range of cytochrome UGT enzymes have been identified in Def-INTESTINAL organoids.

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, CYP2J2 and (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.

(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 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.

 

(A)

Def-INTESTINAL organoids can transport Rhodamine 123

(B)

Immunostaining of intestinal organoids showing localization of the MDR1 transporter protein

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.

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info@definigen.com