Twenty‐four‐well Transwell chamber inserts (Corning, Corning, NY, USA) and Corning BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Corning) were used in migration and invasion assays, respectively; 2 × 104 cells in 200 μL of serum‐free medium were seeded into the upper chamber, and 750 μL medium supplemented with 10% FBS was added into the lower chamber. The cells were incubated for 24 hours and then fixed with 4% paraformaldehyde for 30 minutes. The cells on the top of the chamber were washed with PBS and then scraped with a cotton swab. The cells adherent to the lower chamber were stained with 0.1% crystal violet for 20 minutes at room temperature and counted in 5 random visible fields under a microscope.
Biocoat growth factor reduced matrigel invasion chamber
Manufactured by Corning
Sourced in United States
The BioCoat Growth Factor Reduced Matrigel Invasion Chambers are a specialized laboratory tool designed for cell invasion assays. They provide a standardized in vitro system to assess the ability of cells to migrate through a reconstituted basement membrane. The chambers contain a pre-coated, growth factor-reduced Matrigel matrix that serves as a barrier to cell invasion.
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Lab products found in correlation
Transwell insert chamber, by Corning (3 mentions)
Cell culture insert, by Avantor (2 mentions)
Harleco hemacolor stain set, by Merck Group (2 mentions)
Crystal violet, by Merck Group (1 mentions)
Chamber insert, by Corning (1 mentions)
24 well transwell chamber inserts, by Corning (1 mentions)
Costar transwell insert, by Corning (1 mentions)
Ht90132, by Merck Group (1 mentions)
Evos xl core imaging system, by Thermo Fisher Scientific (1 mentions)
Eosin y solution, by Merck Group (1 mentions)
Dmi1 inverted microscope, by Leica (1 mentions)
Stemi 508 stereo microscope, by Zeiss (1 mentions)
Glycergel mounting medium, by Agilent Technologies (1 mentions)
Eclipse 80i microscope, by Nikon (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Biocoat control insert, by BD (1 mentions)
Dmla microscope, by Leica (1 mentions)
24 protocols using biocoat growth factor reduced matrigel invasion chamber
1
Transwell-Based Cell Migration and Invasion Assay
2
Cell Migration and Invasion Assay
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Transwell assays were applied with chamber inserts (Corning, NY, USA) and Corning BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Corning) for migration and invasion assay, respectively. Lower chamber was filled with 750 μL medium containing 10% FBS, and 3 × 104 cells in 200 μL serum-free medium were added in the upper chamber. After 24 hours incubation, cells on the bottom of the lower chambers were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet (Sigma‐Aldrich). Then the chambers were gently washed with PBS, scraped by cotton swabs to remove the cells on the top, and then finally counted under a microscope in 5 random fields of view.
3
Gastric Cancer Cell Migration and Invasion
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Cell migration and invasion potencies were studied by wound-healing and Transwell. In wound-healing assay, gastric cancer cells were seeded into 6-well plates and grow reach to 100% confluence, the single layer cells were scratched straightly with pipette tip. At 0 hour and 24 hours the distance covered by migrated cells was identified. 24-well Transwell chamber inserts (Corning, Corning, NY, USA) and Corning BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Corning) were used respectively in migration and invasion assays. 6 × 104 HGC 27 and 4 × 104 MGC 803 were resuspended in 200 μL RPMI 1640 medium and laid into upper chamber, meanwhile 750 μL RPMI 1640 with 10% FBS was added into lower chamber. The plate was incubated for 24 hours at 37 °C in humidified incubator containing 5% CO2. Afterwards, medium was discarded, cells were fixed with paraformaldehyde for 15 minutes and then stained with crystal violet for 15 minutes. Random fields were captured under microscope at 100×.
4
Measuring Cell Migration and Invasion
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Cell migration and invasion capabilities were measured using Transwell chamber inserts (Corning, NY, USA) and Corning BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Corning, NY, USA), respectively. 750 μl DMEM with 10% FBS was added into the lower chamber, 3 × 104 cells with 200 μl serum-free medium were plated in the upper chamber. After incubation for 24 hours, cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. Cells remaining on the upper membrane were gently washed with PBS and scraped by cotton swabs. Stained cells on the bottom of the chamber were counted under a microscope on 5 random fields.
5
Cell Migration and Invasion Assay
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Transwell chamber inserts (Corning, NY, USA) and Corning BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Corning, NY, USA) were used for the migration and invasion assays. HGC-27 (2 × 104 cells per well), MGC803 (3 × 104 cells per well), BGC823 (8 × 104 cells per well), and AGS (10 × 104 cells per well) were resuspended in 200 μL of medium without FBS and then placed in the upper chamber. The substrates were placed in a 24-well plate filled with 600 μL of complete medium. The plates were incubated in a humidified incubator at 37 °C containing 5% CO2 for 24 h. After incubation, the cells were fixed in 4% paraformaldehyde for 30 min and then stained with crystal violet for 30 min. Five random fields were selected under an inverted microscope at ×100 magnification, where images were taken and the cell number was counted.
6
Cell Migration and Invasion Assay
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Cell migration and invasion capabilities were measured using Transwell chamber inserts (Corning, NY, USA) and Corning BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Corning, NY, USA), respectively. 750 μl DMEM with 10% FBS was added into the lower chamber, 3 × 10 4 cells with 200 μl serum-free medium were plated in the upper chamber. After incubation for 24 hours, cells were xed with 4% paraformaldehyde and stained with 0.1% crystal violet. Cells remaining on the upper membrane were gently washed with PBS and scraped by cotton swabs. Stained cells on the bottom of the chamber were counted under a microscope on 5 random elds.
7
Breast Cancer Cell Invasion and Migration Assay
8
Melanoma Cell Invasion Assay
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Invasion assays were performed in BioCoat Growth Factor Reduced Matrigel Invasion Chambers (Cat #354483, Corning, Corning, NY, USA) using melanoma cells expressing the indicated shRNAs and melanoma samples treated for 24 h with XL413. The cells were serum-starved for 6 h, and 5×104 cells/insert were seeded in triplicate in the top chamber containing low-serum medium. The cells were then incubated for 20 h to allow invasion toward the serum-rich medium in the bottom well. The number of cells invading the Matrigel was quantified by DAPI staining and imaging; 8–12 fields per membrane were counted, and nuclei quantification was performed using ImageJ software.
9
Cell Invasion Assay with Matrigel
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Cells were treated with 10 μg/mL mitomycin C for 2 hours and then split with trypsin/EDTA. A total of 5 × 104 cells were seeded on top of 8 μm chambers without (cell culture inserts; catalog 10769-242, VWR) and with Matrigel (BioCoat Growth Factor Reduced Matrigel Invasion Chambers; catalog 354483, Corning) and then grown with 0.1% FBS-containing medium on top and 10% FBS-containing medium on the bottom (81 (link)). After 48 hours, cells were removed from the top of the filter inserts, and cells that had passed through the 8 μm pore membranes were fixed with methanol and revealed with the Harleco Hemacolor Stain Set (MilliporeSigma). Images were captured by microscopy and quantitation was performed by counting cells in 3 random fields at ×10 original magnification.
10
Invasion Assay Using Matrigel Chambers
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Four hours after transfection, normal (10% FBS) cell medium was replaced with 2% FBS media. The following day, Corning BioCoat growth factor reduced Matrigel invasion chambers (Corning; New York, USA) were warmed and rehydrated. Initial experiments evaluated optimum cell density for each line (data not shown), and invasion assays were carried out as we have described previously (34 (link)), before Eosin Y solution (Sigma-Aldrich) staining and being photographed on an EVOS XL Core Imaging System (ThermoFisher Scientific).
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