Boyden Transwell migration chambers, 8 µm (Costar) were left uncoated and used to assess chemotactic migration. Cells from stable lines, or uninfected controls, were serum starved for 8–20 h, lifted, counted and 50,000 cells per well, in serum free media, were added to the top of the transwell chamber. Cells were serum-starved for an additional 3 h and then DMEM/EGF (25 ng/ml) or GABA (50 µM) was added to the bottom well and cells were allowed to migrate for 12 h. The bottom of the membrane was fixed in 4% PFA, stained with DAPI and mounted on coverslips. 10 fields of cells on the membrane were counted, per experiment, using fluorescence microscopy. The data are presented as the number of MDA-MB-231 cells that migrated to the bottom membrane.
Boyden transwell migration chambers
Manufactured by Corning
The Boyden Transwell migration chambers are a tool used for studying cell migration and invasion in vitro. The chambers consist of an upper and lower compartment separated by a porous membrane. Cells are seeded in the upper compartment and their migration through the membrane to the lower compartment can be quantified, providing insights into cellular behavior and responses.
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2 protocols using boyden transwell migration chambers
1
Transwell Assay for Chemotactic Migration
2
Quantifying Cell Migration via Transwell Assay
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Boyden Transwell migration chambers (Costar) were coated with 20 μg/ml fibronectin on the bottom chamber. Transfected cells were counted, and 20,000 cells/well in serum-free media were added to the top well. The lower wells were filled with DMEM/10% FBS, and cells were allowed to migrate for 6 h. The top and bottom of the membrane was fixed in 4% paraformaldehyde/PBS, stained with DAPI, and mounted on coverslips. Ten fields of cells on the membrane were counted per experiment using fluorescence microscopy. The data are presented as number of transfected cells that migrated to the bottom membrane divided by number of cells that remained on the top membrane.
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