Measurement of endothelial cell migration using an improved linear migration assay. Presented at the 1995 Microcirculatory Society Meeting.
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.
Hoying
J B
JB
Department of Surgery, University of Arizona Health Sciences Center, Tucson 85724, USA.
Williams
S K
SK
D000818:Animals; D002196:Capillaries / Q000166:cytology; D002452:Cell Count / Q000295:instrumentation / Q000379:methods*; D002465:Cell Movement; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology*; D005353:Fibronectins; D005456:Fluorescent Dyes; D006801:Humans; D007091:Image Processing, Computer-Assisted; D012685:Sepharose; D013194:Staining and Labeling; D013269:Stochastic Processes
D005353:Fibronectins; D005456:Fluorescent Dyes; D012685:Sepharose