Migration and invasion assays were performed using uncoated and Matrigel-coated Transwell (Corning Costar Corp., Cambridge, MA) membrane filter inserts with pore size of 8 μm according to the manufacturer’s instructions. Approximately 2×104 cells per well were suspended in 200 μL medium with 1% FBS and then seeded into the upper chambers filled with 500 μL medium containing 10% FBS. The cells were incubated at 37°C for 12 h and 24h for migration and invasion respectively. The cells on the upper surfaces of the Transwell chambers were removed by cotton swabs;the cells on the lower surface of the membrane were fixed and stained with crystal violet solution. The number of cells was counted in five randomly selected microscope fields.
Matrigel coated transwell
Manufactured by Corning
Sourced in United States
The Matrigel-coated Transwell is a laboratory equipment designed for cell culture applications. It consists of a porous membrane that separates two compartments, allowing the study of cell migration and invasion through the membrane. The membrane is coated with Matrigel, a basement membrane extract, providing a more physiologically relevant substrate for cell-based assays.
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Lab products found in correlation
Transwell insert, by Corning (3 mentions)
Axio observer a1, by Zeiss (3 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions)
Hepatocyte growth factor (hgf), by Thermo Fisher Scientific (2 mentions)
Eclipse ci l, by Nikon (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Chopstick electrode, by World Precision Instruments (1 mentions)
Cellsens dimension 1, by Olympus (1 mentions)
Bx63 microscope, by Olympus (1 mentions)
Dp80 camera, by Olympus (1 mentions)
Cck 8 reagent, by Merck Group (1 mentions)
Brilliant blue r 250, by Merck Group (1 mentions)
Eclipse te2000, by Nikon (1 mentions)
Crystal violet, by Wuhan Servicebio Technology (1 mentions)
Uncoated transwells, by BD (1 mentions)
Dexamethasone (dex), by Thermo Fisher Scientific (1 mentions)
Stemdiff definitive endoderm kit, by STEMCELL (1 mentions)
Cts knockout dmem f12, by Thermo Fisher Scientific (1 mentions)
26 protocols using matrigel coated transwell
1
Transwell Invasion and Migration Assay
2
Wound Healing and Cell Metastasis Assays
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For wound-healing, cells were seeded in the 6-well plate and cultured for 8–24 h. When cells grew to 90% confluence, each well with cultured cells was scratched by a 200 μl pipette tip followed by 48 h culture. The photos of cultured cells were captured at 0, and 48 h in the same position and the area of wound-healing was analyzed.
Transwell assay was applied to evaluate the changes in cell metastasis ability. Cells were seeded in the Matrigel-coated Transwell or normal Transwell plate (Costar, USA) of the upper chamber at 5 × 104 cells/well. The bottom of the Transwell plate was added with 400 μL DMEM with 10% PBS. Following 24 h incubation, cells on the upper surface of the Transwell filter membrane were wiped off, and the membrane and lower chamber cultures were stained with 0.1% crystal violet. After dried, the staining cells were observed at three random fields under a microscope.
Transwell assay was applied to evaluate the changes in cell metastasis ability. Cells were seeded in the Matrigel-coated Transwell or normal Transwell plate (Costar, USA) of the upper chamber at 5 × 104 cells/well. The bottom of the Transwell plate was added with 400 μL DMEM with 10% PBS. Following 24 h incubation, cells on the upper surface of the Transwell filter membrane were wiped off, and the membrane and lower chamber cultures were stained with 0.1% crystal violet. After dried, the staining cells were observed at three random fields under a microscope.
3
Angiogenic Potential of Irradiated HUVECs
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Irradiated HUVECs were re-seeded onto Matrigel-coated transwell (Corning, NY, USA), followed by treatment with or without CA for 6 h. Total tube length was observed under a light microscope and plotted using Image J.
4
Assessing Melanoma Cell Migration and Invasion
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The migration and invasion abilities of melanoma cells were assayed using Transwell inserts and Matrigel-coated Transwell (Corning, MA, USA). Briefly, 2 × 104 melanoma cells were suspended in 200 μL of medium free of serum, and added into the top chamber. 500 mL culture medium containing 10% FBS was added into the lower chamber. 24 h after incubation, a cotton swab was used to remove cells on the upper surface of the membrane gently and the cells were fixed in 4% paraformaldehyde, then 0.5% crystal violet was used to stain these cells for 20 min. The migrating or invading cells were photographed and counted under a microscope.
5
Transwell-based Cell Migration and Invasion Assay
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For the migration and invasion assays, cells were seeded in the upper chamber of a transwell (8-μm pore size) or in a Matrigel-coated transwell (Corning, NY, USA) in serum-free media. The lower chamber contained 600 μl of basal medium with 10% fetal bovine serum as a chemoattractant. After 48 h of incubation, the nonmigrated or noninvaded cells were gently removed from the upper chamber using a cotton swab. The remaining cells in the lower chamber were fixed with methyl alcohol, stained with a 0.1% crystal violet solution for 15 min and then imaged via microscopy (Olympus Corporation, Tokyo, Japan). For each sample, five random fields (×100 magnification) were selected, and the cells in each field of view were counted.
6
Transwell Migration and Invasion Assay
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Approximately 2 × 104 cells in 300 μL DMEM medium without FBS were seeded into upper transwell chamber (8 μm pore size) to evaluate cell migration. The lower chamber was filled with 800 μL DMEM medium supplemented with 10% FBS. After 24 h, the cells attached to the lower surface of the membrane were fixed with 4% formaldehyde, stained with 0.5% crystal violet, and then counted under a microscope in five random fields. Invasion assays were performed under the same conditions as the migration assays, but in matrigel-coated transwell (Corning, NY, USA) inserts.
7
Cell Migration and Invasion Assays
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Breast cancer cells (5 × 104) treated with siRNAs (siCtrl or siNEK8) were seeded in a Transwell plate with serum-free culture medium (Corning, Corning, NY, USA). The migrated cells were fixed with 4% paraformaldehyde, stained using crystal violet, and photographed under an inverted microscope (Nikon Eclipse Ci-L, Tokyo, Japan).
Invasion assays were performed in the same manner using a Matrigel-coated Transwell (Corning, Corning, NY, USA). In five random fields, the cells that were stained were counted, and the average number was determined.
Invasion assays were performed in the same manner using a Matrigel-coated Transwell (Corning, Corning, NY, USA). In five random fields, the cells that were stained were counted, and the average number was determined.
8
RA08 Inhibits Cell Invasion Assay
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1 × 105 cells resuspended in serum-free medium were seeded in the upper chamber of a Matrigel-coated Transwell (Corning), and the lower chamber was filled with serum-supplemented medium. Cells were incubated at 37 °C for 24 h and fixed with 4% paraformaldehyde (Sigma-Aldrich). After removal of the Matrigel, the cells that migrated to the lower surface of the membrane were stained with Hoechst-33342 (5 ng/ml, Sigma-Aldrich) and were counted under the microscope. Three independent wells were counted for each condition. To assess RA08’s effects on cell invasion (and RGD peptide), cells were previously treated for 48 h with different concentrations (0 nM, 100 nM, 200 nM, 500 nM and 1000 nM), which were also present in the medium during the cell invasion assay.
9
Evaluating Migration and Invasion Abilities
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The migration and invasion abilities of GC cells were assayed using Transwell inserts and Matrigel-coated Transwell (Corning, MA, USA). In brief, the CM collected from CAFs or NFs was added into the lower chamber of a 24-well plate. MGC-803 and SGC-7901 cells, with a density of 2 × 104 cells in 200 μL of serum-free medium, were added into the Transwell chambers with the Matrigel member covered or uncovered. Then, the cell plate was incubated in a humidified atmosphere (37 °C and 5% CO2). After culturing for 24 h (migration assays) or 48 h (invasion assay), the cells on the upper surface of the membrane were completely removed using a cotton swab. Then, the cells were fixed in 4% paraformaldehyde and stained with crystal violet solution for 15–20 min. The migrating or invading cells were counted under a microscope and photographed.
10
Transwell Invasion Assay
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Stabilization pool cells were cultured in serum‐free medium in the upper chamber of a Matrigel‐coated Transwell (Corning, NY) to determine cell invasion. The lower chamber contained a standard medium with 10% fetal bovine serum. Cells migrating to the submembrane surface were fixed with formalin, stained with 0.05% crystal violet, counted, and imaged under a microscope after 24 h of incubation.
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