A modified two-chamber migration assay (8-mm pore size, BD Biosciences, Bedford, MA) was adopted to assess cell mobility according to the manufacturer's instructions as described previously 20 (link). Approximately 2 × 104 cells were plated into the upper chamber, and the cells were then allowed to migrate into the lower chamber for 18-24 hours. The cells at the bottom of the membrane were fixed and stained with methanol 20%/crystal violet 0.2%, while the cells in the upper chamber were removed using cotton swabs. The data are presented as the means ±standard error of the mean (SEM).
Modified two chamber migration assay
Manufactured by BD
Sourced in United States
The modified two-chamber migration assay is a laboratory equipment designed to measure the migratory capabilities of cells. It consists of two interconnected chambers separated by a porous membrane, allowing the study of cell movement and behavior in response to various stimuli.
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2 protocols using modified two chamber migration assay
1
Cell Migration Assay using Two-Chamber System
2
Modified Transwell Assay for HepG2 Cell Migration and Invasion
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Migration of HepG2 cells was assessed using a modified two-chamber migration
assay (BD Pharmingen, San Diego, CA, USA) with a pore size of 8 mm. After 50 μM
kaempferol treatment and/or miR-21 mimic transfection, 1 × 103 HepG2
cells were suspended in 200 mL serum-free DMEM and seeded into top chamber.
Complete DMEM (600 mL) was added into the lower chamber. After incubation for
48 h, cells were fixed with methanol (Beyotime Biotechnology, Shanghai, China)
immediately. Non-traversed cells in top chamber were moved using cotton swab
carefully and traversed cells in lower chamber were counted under microscope
(Nikon, Japan). Cell migration (%) was calculated as follows: average traversed
cells in treatment (transfection) group/average traversed cells in control group
× 100%.
Cell invasion was conducted similarly with the cell migration assay except that
the polycarbonate filter was pre-coated with Matrigel (BD Biosciences, Franklin
Lakes, NJ, USA) in line with the manufacturer’s instruction.
assay (BD Pharmingen, San Diego, CA, USA) with a pore size of 8 mm. After 50 μM
kaempferol treatment and/or miR-21 mimic transfection, 1 × 103 HepG2
cells were suspended in 200 mL serum-free DMEM and seeded into top chamber.
Complete DMEM (600 mL) was added into the lower chamber. After incubation for
48 h, cells were fixed with methanol (Beyotime Biotechnology, Shanghai, China)
immediately. Non-traversed cells in top chamber were moved using cotton swab
carefully and traversed cells in lower chamber were counted under microscope
(Nikon, Japan). Cell migration (%) was calculated as follows: average traversed
cells in treatment (transfection) group/average traversed cells in control group
× 100%.
Cell invasion was conducted similarly with the cell migration assay except that
the polycarbonate filter was pre-coated with Matrigel (BD Biosciences, Franklin
Lakes, NJ, USA) in line with the manufacturer’s instruction.
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