Transfected cells were collected and suspended to a density of 1 × 105 cells/ml. For cell migration assay, upper chamber was equipped with 8-µm porous membranes (BD Biosciences) and added with 100 µl of the suspension. Six hundred µl of complete culture medium was added to each lower chamber to induce migration. The non-migrated cells were removed with a cotton swab after incubation for 24 h at 37°C with 5% CO2 whereas the migrated cells were fixed with 4% (v/v) paraformaldehyde for 20 min and stained for 20 min with 0.1% crystal violet. The number of migrated cells was taken by an inverted microscope. For the cell invasion assay, chambers were precoated with Matrigel (BD Biosciences).
8 m porous membranes
Manufactured by BD
The 8-µm porous membranes are a laboratory filtration product designed for separating and filtering materials. The membranes have a pore size of 8 micrometers, which allows for the efficient filtration of various substances. This product is intended for use in controlled laboratory settings and environments.
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2 protocols using 8 m porous membranes
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Cell Migration and Invasion Assay
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Cell Migration and Invasion Assay Protocol
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Transfected cells were trypsinized, washed, centrifuged, and collected. The cells were mixed with FBS-free culture medium to yield a cell suspension at a density of 1×105 cells/ml. Regarding cell migration assay, 100 µl of the suspension was added to each upper chamber of wells equipped with 8-µm porous membranes (BD Biosciences), whereas 600 µl of complete culture medium was added to each lower chamber to induce migration. After 24 h of incubation at 37°C in an incubator supplied with 5% CO2, the cells that had not migrated to the lower chamber were removed with a cotton swab, whereas the migrated cells were fixed for 20 min with 4% (v/v) paraformaldehyde and stained for 20 min with 0.1% crystal violet. The number of migrated cells was counted in five randomly selected fields under an inverted microscope (magnification, ×100; Olympus Corp.). Regarding the cell invasion assay, chambers were precoated with Matrigel (BD Biosciences). All subsequent steps were performed as described for the cell migration assay.
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