For migration, the Transwell cell culture chambers (Corning Costar Corp, Cambridge, MA, USA) were used. The two transfected cells were seeded into the upper chamber with serum-free medium, while the bottom chamber was filled with 600 ml 10% FBS medium. For invasion, the BioCoat™ Matrigel Invasion Chamber 24-Well Plate 8.0 Micron (Corning, NY, USA) was used. The two transfected cells were seeded into the upper chamber with serum-free medium, while the bottom chamber was filled with 600 ml 20% FBS medium. The MDA-MB-231 plates were incubated at 37 °C with 5% CO2, and the MDA-MB-468 plates were incubated at 37 °C with no CO2. After 24 h, the indicated cells were fixed with 4% PFA (Sigma) for 30 min, washed with PBS, then stained with 0.01% crystal violet for 20 min, and then finally air dried.
Biocoat matrigel invasion chamber 24 well plate 8.0 μm
Manufactured by Corning
Sourced in United States
The BioCoat Matrigel Invasion Chamber 24-well plate 8.0 μm is a laboratory equipment product designed for cell invasion assays. The plate contains a layer of Matrigel matrix that serves as a barrier to simulate the extracellular matrix, allowing the evaluation of a cell's ability to invade through this barrier.
Automatically generated - may contain errors
5 protocols using biocoat matrigel invasion chamber 24 well plate 8.0 μm
1
Transwell Migration and Invasion Assay
2
Transwell Assay for Cell Migration and Invasion
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For migration, transwell cell culture chambers (Corning Costar Corp, Cambridge, MA, USA) were used. The three transfected cell types (TPC1 3 × 10 4 cells; BCPAP 5 × 10 4 cells; FTC133 4 × 10 4 cells) were seeded into the upper chamber with serum-free medium, and the bottom chamber was filled with 600 ml 10% FBS medium. For invasion, BioCoat™ Matrigel Invasion chamber 24-Well Plate 8.0 Micron (Corning) was used. Cells treated with siRNA or plasmid vector (8 × 10 4 cells) were seeded onto inserts in serum-free medium, and the inserts were placed into a 24-well plate filled with 20% FBS medium. Following incubation for 24 h, cells in the bottom chamber were fixed with 4% PFA (Sigma), stained with 0.01% crystal violet, imaged under a microscope at a magnification of 200× and quantified with the ImageJ software (https://imagej.nih. gov/ij/). All experiments were replicated three times.
3
Cell Migration and Invasion Assay
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Cells were transfected with siRNAs as described above. Transwell chambers were used for cell migration (BioCoat Control Insert 24-well plate 8.0 μm; Corning Inc., Corning, NY, USA) and invasion (BioCoat Matrigel Invasion Chamber 24-well plate 8.0 μm; Corning Inc.) analyses. Cells were harvested 24 h after transfection and resuspended in serum-free RPMI 1640 medium, after which 5 × 104 cells were added to the upper chamber. RPMI 1640 medium with 10% fetal bovine serum was added to the lower well. After incubation for 24 h at 37 °C, migrating or invading cells on the lower surface of the filter were fixed and stained using a Diff-Quik staining kit (Sysmex, Tokyo, Japan). Cell numbers were determined by counting in five randomly selected microscope fields per membrane.
4
Cell Migration and Invasion Assay
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Cells were transfected with plasmids as described above. Transwell chambers were used for cell migration (BioCoat Control Insert 24-well plate 8.0 μm; Corning Inc., Corning, NY, USA) and invasion analyses (BioCoat Matrigel Invasion Chamber 24-well plate 8.0 μm; Corning Inc.). Cells were harvested 24 h after transfection and resuspended in culture medium containing 1 mg/ml bovine serum albumin, after which 1 × 105 cells were added to the upper chamber. Culture medium with 10% fetal bovine serum was added to the lower well. After incubation for 24 h at 37°C, migrating or invading cells on the lower surface of the filter were fixed and stained using a Diff-Quik staining kit (Sysmex, Tokyo, Japan). Cell numbers were determined microscopically by counting in five randomly selected fields per membrane.
5
Quantifying cell migration and invasion
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Cell migration and invasion assays were performed as described previously8 (link). Briefly, OSCC cells were transfected with siRNAs as described above and incubated for 48 h. Transwell chambers were used for cell migration (BioCoat Control Insert 24-well plate 8.0 μm; Corning Inc., Corning, NY, USA) and invasion (BioCoat Matrigel Invasion Chamber 24-well plate 8.0 μm; Corning Inc.) analyses. The numbers of migrated or invaded cells were determined microscopically by counting in eight randomly selected microscopic fields per membrane.
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