Matrigel invasion system

The migration activity of HCAECs was determined using the growth factor-reduced Matrigel invasion system (Becton Dickinson) as previously described.[19 (link)] HCAECs (5 × 10⁴ cells) were seeded on top of the ECMatrix gel (Chemicon International, Inc., Temecula, CA) and incubated at 37°C for 4 h during hypoxia with addition of chemerin, TNF-alpha, PD98059, an anti-TNF-alpha antibody, chemerin siRNA, or scrambled siRNA. Three different phase-contrast microscopic high-power fields per well were photographed. The migratory HCAECs were stained with Liu stain (BaSO, New Taipei City, Taiwan) and counted. The observer was blind to the experiment.

We determined the migration activity of HCASMCs using the growth factor-reduced Matrigel invasion system (Becton Dickinson, Franklin Lakes, NJ) following manufacturer’s protocol. We then seeded the Matrix gel (Chemicon International, Inc., Temecula, CA) with 5 × 10⁴ cells. Cells were then incubated at 37°C for 4 hours, with or without hypoxia. Three different phase-contrast microscopic high-power fields per well were photographed. The migratory cells with positive stain were counted by an observer blind to the experiment.