Transwell Invasion Assay

Overview

The transwell invasion assay (Boyden chamber assay) measures the ability of cancer cells to invade through an extracellular matrix barrier — typically Matrigel-coated membranes — toward a chemoattractant in the lower chamber. Invaded cells are stained and counted after a fixed incubation period. It is a standard in vitro surrogate for invasive capacity and epithelial-mesenchymal transition (EMT).

Used by

• Used in GBC cell lines (GBC-SD, NOZ) to measure invasive capacity; conditioned medium from stiff-matrix-activated gallbladder fibroblasts increased invasion, an effect abrogated by SEMA7A knockdown in fibroblasts; recombinant human SEMA7A rescued invasion via ITGB1 binding (RGD motif dependent) and was blocked by integrin β1 neutralizing antibody or RGD peptide PMID:24997986

Notes

• Commonly 8 µm pore Transwell inserts coated with Matrigel for invasion (vs uncoated for migration only).
• Results are typically normalized to a control condition and expressed as fold-change in invaded cell number.
• Often paired with tumorsphere assays to jointly assess EMT and stemness.

Sources

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