Cell-based Assays for Drug Transport
Millicell-96 cell culture insert plate is a patented
96-well device designed to support epithelial cell growth and differentiation
of Caco-2 and other cell lines.
After the formation of a
differentiated cell monolayer, the device can be used to measure the rate of
drug transport across the cell barrier. All procedures are designed to be
carried out in a single device. The assay can be performed using automation for
cell seeding, cell feeding, washing and drug transport experiments.
The Millicell-96 device consists of a filter plate with 96 basolateral access
holes, growth plate (single well and 96-well available), lid and 96-well
receiver plate.
- ANSI/SBS-compliant design – the
Millicell-96 device is compatible with automated liquid handling systems and
span pipettes
- Results not compromised – ease of access to both apical
and basolateral sides is assured
- Increased sample throughput – using
the 96-well system, throughput is increased by four compared to 24-well systems
- Versatile format – 96- and single-well feeder trays available
Performance
Drug Permeability Rates Correlate to Human Absorption
)
Drug compounds representing active, passive and efflux transporters were tested at ArQule, Inc. using the Millicell-96 assay system (for a complete list of compounds, refer to Millipore Application Note AN1727EN00). Each compound’s permeability rate was plotted against their percent human absorption values. A nonlinear regression curve was generated using Statistica™ software. The resultant curve provided a good fit to all of the data points to demonstrate that the Millicell-96 assay system could correctly identify and classify the permeability properties of all 25 compounds tested.
Drug Permeability Rates Correlate with 24-well Assay Systems
)
Caco-2 drug transport rates using 10 tritium labeled drugs were measured using the Millicell-96 assay system and compared to the drug transport rates obtained from an established 24-well system. This set compares the results from four separate experiments. The R2 value of 0.99 suggests that the data between the two formats (24-well assay vs. Millipore 96-well) correlate. For a complete listing of the drugs tested, refer to Millipore Application Note AN1727EN00.
Reproducible Assay Results
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Low variability of in vitro cell-based assay platforms is critical for reliable assay results. This figure examines the reproducibility of drug transport rates in Millicell-96 assay system using a 21-day culture of Caco-2 cells and a single drug, propranolol, in all 96-wells of every plate tested. Transport rates were determined in 3 experimental runs performed on different days, each run consisting of 4 different production lots of Millicell-96 plates. Analysis of variance on the plate average, inter-plate and intra-plate standard deviations does not vary significantly among the production lot tests or days. The total standard deviation is calculated to be 0.3 x 10-5 cm/sec.
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