|Incubate at 37 C in CO2 incubator; incubation times may be varied 48hours.
The in vitro invasibility was assessed using the CultreCoat® 96-Well BME-Coated Cell Invasion Optimization Assay Kit supplied by R & D Systems. The cells starving in serum-free medium for 16 h were seeded at 25000 cells/well in the upper chamber of Transwell, in the meantime, 2.5 and 5 mg/ml amygdalin was added to continue the culture for 48 h. Then, the number of invasion cells was analyzed in accordance with the kit instructions. The wound-healing assay was used to assess the in vitro migratability of cancer cells. Specifically, the cells were cultured in 6-well plate until single-layer confluence. After starving in serum-free medium overnight, the 200 μ L pipette tip was used to make scorings in the cell layer. Then, 2.5 and 5 mg/ml amygdalin was added to continue the culture for 48 h. The distance between the scorings was observed and measured. Full paper
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