Quantitative high throughput endothelial cell migration and invasion assay system.
We have developed a novel assay system combining fluorescent signal-blocking microporous PET membrane inserts and ECM to study dynamic endothelial cell migration and invasion. The assay described here may be applicable to other cell types for analogous assay. The currently used method for analyzing migration and invasion requires the removal of nonmigratory cells from the top of a clear microporous membrane. This laborious step is required to quantitate the invaded or migrated cells on the bottom of the membrane. When the fluorescent signal blocking the microporous membrane is used, the fluorescent signal from noninvaded or nonmigratory cells from the top of the membrane is virtually eliminated and allows for rapid and efficient measurement of the migrated or invaded cells on the underside of the insert. Unlike conventional low throughput migration assays, this assay is easy to perform and provides a quantitative invasion/migration profile of the chemoattractant of choice. This robust automation compatible assay will serve as a powerful tool to screen potential antiangiogenic compounds for drug discovery.
James C
JC
Maliakal
D016475:3T3 Cells; D043168:Angiostatins; D000818:Animals; D002465:Cell Movement; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D008222:Lymphokines / Q000494:pharmacology; D051379:Mice; D009361:Neoplasm Invasiveness; D010446:Peptide Fragments / Q000494:pharmacology; D010958:Plasminogen / Q000494:pharmacology; D011509:Proteoglycans / Q000494:pharmacology; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D004338:Drug Combinations; D016228:Endothelial Growth Factors; D007797:Laminin; D008222:Lymphokines; D010446:Peptide Fragments; D011509:Proteoglycans; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; C059485:matrigel; D043168:Angiostatins; D010958:Plasminogen; D003094:Collagen