Cell migration and invasion were assessed using BD BioCoat Matrigel Invasion Chambers and Control Inserts (BD Biosciences, San Jose, CA, USA). The same number cells were plated in 6cm plates with a growth medium cultured for 24 hours. The medium was removed and erlotinib and/or chelerythrine were applied to treat the cells for 24 hours. The viable cells were collected and seeded on either control inserts (a polyethylene terephthalate membrane) or trans-well chambers. A 2ml medium supplemented with 15% FBS was added to the lower chamber, which served as the chemoattractant. 4×104 cells were re-suspended in the media plus 1% FBS was added to the upper chamber (0.5ml). 20 hours later, migrating or invading cells attached to the lower surface of the membrane insert which were fixed and stained and then counted under a microscope [16 ]. Relative migration was calculated by comparing cells transfected with the negative control. The percentage invasion was calculated based on the number of cells which had invaded through the Matrigel insert, divided by the number of cells which had migrated through the control insert.
Biocoat matrigel invasion chambers and control inserts
Manufactured by BD
Sourced in United States
The BD BioCoat Matrigel Invasion Chambers and Control Inserts are laboratory equipment designed for cell invasion assays. The chambers are pre-coated with a layer of Matrigel basement membrane matrix, which serves as a barrier to simulate the extracellular matrix. The control inserts do not contain the Matrigel coating. These products allow for the assessment of a cell's ability to invade through the matrix.
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5 protocols using biocoat matrigel invasion chambers and control inserts
1
Cell Migration and Invasion Assay
2
Cell Migration and Invasion Assays
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Cell migration and invasion assays were conducted using BD BioCoat Matrigel Invasion Chambers and Control Inserts (BD Biosciences; Becton, Dickinson and Company). A total of 5×104 SC or knockdown of SK-BR-3 cells were re-suspended in 0.5 ml medium with 1% fetal bovine serum (FBS), then seeded on either upper control inserts or trans-well chambers with Matrigel (BD Biosciences; Becton, Dickinson and Company). A total of 2 ml medium containing 15% FBS was added onto the lower chamber, which served as a chemo-attractant. A total of 20 h later, migrating or invading cells that had attached to the lower surface of the membrane insert were fixed at room temperature for 2 min and stained by Giemsa at room temperature for 10 min, then counted under a light microscope (Olympus IX51; Olympus Corporation) at ×200 magnification.
3
Matrigel Invasion Assay for Cell Migration
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Invasion of cells in vitro was assayed using the BD BioCoat Matrigel Invasion Chambers and Control Inserts (BD Biosciences, Bedford, MA, USA) respectively. Each well of a 24-well plate contained an insert with an 8-μm pore size polyethylene terephthalate membrane. Cells (1×105 per Transwell) were suspended in serum-free DMEM and seeded into the upper chamber. DMEM containing 2% fetal bovine serum was then added to the bottom chamber of 24-well plates to serve as a chemoattractant. After 48 h of incubation, cells on the upper surface of the filter were removed, and cells that migrated to the lower surface were fixed and stained with 1% toluidine blue. For quantification of cell invasion, 10 fields per experimental condition were randomly selected as previously described (28 (link)) and micrographed with IX71 microscope (Olympus, Tokyo, Japan). Images are representative of at least three independent experiments.
4
Cell Migration and Invasion Assay
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Invasion and migration assays were performed using the BD BioCoat Matrigel Invasion Chambers and Control Inserts (BD Biosciences), respectively. Briefly, cells (1 × 105 cells/well) were seeded with medium containing low serum (1%) in the upper chamber. The lower chamber was filled with medium containing high serum (20%) as a chemoattractant. Cells were incubated for 48 hrs and then membranes were stained using Diff-Quick (Siemens). A light microscope was used to count the number of migrating and invading cells.
5
Evaluating Cell Migration and Invasion
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Cell migration and invasion were assessed by using BD BioCoat Matrigel Invasion Chambers and Control Inserts (BD Biosciences). The cells were transfected with either anti-miR-675 or SC (40nM), then seeded on either control inserts (polyethylene terephthalate membrane) or trans-well chambers with Matrigel. Two ml RPMI supplemented with 15% FBS was added to the lower chamber, served as the chemo-attractant. 0.5x104 transfected Fadu cells were re-suspended in RPMI plus 1% FBS, added to the upper chamber (0.5 mls). Twenty hours later, migrating or invading cells attached to the lower surface of the membrane insert were fixed and stained, then counted under a microscope. Relative migration was calculated by comparison with cells transfected with the negative control. The percentage invasion was calculated based on the number of cells which have invaded through the Matrigel insert, divided by the number of cells which have migrated through the control insert.
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