MDCK Permeability

MDCK Permeability 2018-05-15T15:19:45+00:00

 

MDCK Permeability

The Madin-Darby canine kidney (MDCK) cell model is one of the commonly used cell monolayer systems to assess the human intestine barrier.

Advantages of MDCK (Madin-Darby Canine Kidney) Cells

  • Easy to use and analytically clean, with good viability and reproducibility
  • Passive diffusional transport information at the cellular level
  • Lack of substantial levels of metabolizing systems or p-glycoprotein pumps
  • Three-day growth period

Assay Methodology

The MDCK permeability screen (uni-directional) is performed to measure test compounds across a monolayer of MDCK cells. The cells seeded on 24- or 96-well plates to form a confluent monolayer are used to determine the efflux of the test compound when added to the apical side of the membrane. The transport of the compound across the monolayer is monitored over a 60 min time period.

MDCK Permeability: Standard Assay Conditions (Customizable)

Testing Compound Concentration
10 μM

Assay Buffer
HBSS, pH 7.4

Plate Format
24-well or 96-well Transwell plates

Transport Period
1 hour at 37C

Detection Method
LC-MS/MS

Control Compounds
Atenolol and propranolol

Deliverables
Papp value for A-B direction

Turnaround Time
5-7 Days

Compound requirements
20 μL 10 mM DMSO stock