Wound-Healing Assay

The wound-healing assay was used to evaluate the migration ability of trophoblasts. Cells in the fresh medium were placed into a 6-well plate chamber (5×10⁵ cells/well) and incubated for 24 h. When the cells reached 80%-90% confluence, a scratch across the cell surface was made by the plastic pipette tip scraped across the cell surface. Phosphate-buffered saline (PBS) was used to remove floating debris, and the wound was photographed immediately (0 h). The cells were then cultured in the serum-free medium. The wounds were photographed at 24 h and 48 h to measure the extent of wound healing. The sizes of the gaps were measured with Image-Pro Plus software.