Abstract
Objective: The under-agarose migration assay developed for use with endothelial cells provides a measurement of the intrinsic migratory behavior of a cell population. However, the assay is labor intensive and lacks experimental flexibility. This migration assay has been refined and tested on human microvessel endothelial cells in the presence of a migration stimulus or on differing matrix-coated substrates.
Methods: The improved assay retains the linear geometry and mathematical basis of the under-agarose assay. Cells migrating from a cell reservoir formed with a Delrin® insert are counted using an automated image-analysis system utilizing a high-contrast, fluorescent nuclear stain. From the cell counts, a stochastic measure of random migration is calculated.
Results: Values for random migration between the improved migration assay and the traditional under-agarose assay were very similar. Furthermore, a stochastic measure of endothelial cell migration on fibronectin was determined.
Conclusions: This improved linear migration assay permits readily obtainable measures of endothelial cell migration for a number of experimental conditions. Improvements in the assay include the use of a removable fence for forming cell reservoirs, a nuclear stain to facilitate cell counting, and a more comprehensive analysis of the cell migration.