Migration and Invasion Assay of Trophoblast Cells

The migration and invasion capacities of HTR-8/SVneo and JAR cells were determined with 8 μm Transwell filters (Corning, USA). The transfected cells were resuspended in serum-free medium and added to the upper chambers of the Transwell. Medium containing 20% FBS was added to the lower chambers. In the migration assay, the cells were incubated for 24 hours at 37°C in 5% CO₂ atmosphere and allowed to migrate to the bottom of the well. These cells were fixed with 4% paraformaldehyde and then stained with crystal violet [25 (link)]. In the invasion test, the upper chamber was coated with diluted Matrigel (BD Biosciences, USA), and after 48 hours of culture, the cells were fixed and stained. Five fields of view were randomly selected for observation under the microscope. The experiment was performed in triplicate [27 (link)].