We have successfully grown HUB Organoids™ as 2D monolayers on Transwells® to analyze tight-junction dynamics and determine barrier function integrity of epithelia in response to various external stimuli, like a drug or chemical compound. Assays results are used to determine your drug EC50/IC50 and can be presented as dose response curves and/or IC/EC values.
Barrier function/integrity is evaluated by measuring the trans epithelial electrical resistance (TEER)
Transport function is assesses by measuring a fluorescent readout such as a PGP probe, across the organoid membrane
Florescent markers such as Lucifer Yellow or Dextran-FITC are measured to evaluate paracellular permeability
Cell viability is determined by quantifying the cellular ATP content
Assay layout: 24- and 96-well plate format
Read out: measurement of fluorescence (permeability/barrier function) and cellular ATP levels (cell viability)