Almost no gap at 0 h — cells were too dense (Wound Healing Assay)

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Accepted Answer

Initial scratch gap is tiny; by 24 h the wound has fully closed, making it impossible to discriminate between groups. Common causes: 1) Seeding density was too high 2) Scratch was not deep/wide enough 3) Cells proliferate too quickly Solutions: 1) Pre-experiment to optimize seeding density 2) Ensure a clearly measurable gap at the 0 h time point 3) If needed, use low-serum medium or add mitomycin C to suppress proliferation