DefiniGEN’s intestinal monolayer product INTReady constitutes a ground-breaking new system to model the permeability of the human intestine and investigate drug efflux. The cells are grown as a monolayer in a similar manner to standard Caco-2 immortalized cell lines with the key benefit of greatly increased biological relevance, as the cells have primary human intestinal cell function, metabolism and genetics.
- Normal human genetics wild-type donor genetics and karyotype verified
- Display multiple key gut markers OLFM4, CHGA, MUC2, Villin and KRT19
- Cells can be used to model the permeability of the human intestine and investigate drug efflux
- Monolayer format grown in a similar manner to standard Caco-2 immortalized cell lines
- Standardized cell product containing human intestinal monolayer cells producing reproducible and biologically relevant data
Confluent monolayer formation has been demonstrated via TEER, (transepithelial electrical resistance), ZO1 (Zonula Occludens/Tight Junction Protein-1) immunostaining, and Lucifer Yellow analysis. TEER measurement of the monolayer demonstrates tight junction formation with typical values of >150ohm x cm2 being observed after 7 days of monolayer culture. These levels are comparable to those observed with a reference Caco-2 line after 20+ days. ZO1 protein localization immunostaining at the cell boundaries of all cells analyzed indicates a confluent monolayer has been formed. Correspondingly, Lucifer Yellow assays to assess the permeability characteristics of the monolayer has established monolayer integrity with observed values of Papp = 0.87 ± 0.32 × 10-6 cm/sec (A to B, 100 μM, 60 min).
Biologically relevant system
Efflux transporter activities
CYP3A4 activity assay
Def-INT monolayer secretes mucus-forming proteins
For validation, a set of compounds were screened with Caco-2 and Def-INT cells grown in monolayer. Permeability assay over a range of permeabilities show a good correlation with Caco-2. Def-INT grown monolayer shows better predictability for Furosemide than Caco-2.