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Transwell migration and invasion assays

JG Jiuping Gao
TY Ting Yang
XW Xu Wang
YZ Yi Zhang
JW Jing Wang
BZ Beilei Zhang
DT Dihong Tang
YL Yanqiong Liu
TG Ting Gao
QL Qiuhui Lin
JT Jun Tang
JC Jingting Cai
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Cell migration assay was performed in a 24-well plate using Transwell inserts (Corning Inc., Corning, NY, USA) with 8.0-µm pore size. The primary spheres and spheres at 4 passage were collected and resuspended in DMEM and CSC-M, respectively. Then, 200 µL of growth factor-free CSC-M containing 5 × 104 cells was loaded onto the upper chamber of the transwell insert, and 600 µL of CSC-M was loaded onto the lower chamber. Cells were allowed to migrate for 24 h at 37 °C. The inserts were washed with PBS, and non-migratory cells that remained in the upper compartment were removed with a cotton swab. The migratory cells in the lower chamber were fixed with methanol and stained with crystal violet (Hengxing Chemical Reagent Co., Ltd., Tianjin, China), and counted using an optical microscope (magnification, x400; Olympus, Tokyo, Japan). Cell invasion assay was performed using matrigel-precoated (Becton, Dickinson and Company, Franklin Lakes, NJ, USA) transwell chambers (8-µm pore size; Corning Inc., Corning, NY, USA) in a 24-well plate.

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