Cell migration assays were performed using modified 24-well Boyden chambers (Costar, Acton, MA, USA), containing a polycarbonate membrane with 8.0-μm pores. HUVECs were starved in basic EGM-2 (serum/growth factor free) overnight at 37°C, prior to being harvested with trypsin and resuspended in basic EGM-2. The single cell suspensions with 20 μM DHA or 20 μM SB203850 were seeded at 1×105 cells/well in the upper chamber, while 0.5 ml EGM-2 with 20 ng/ml VEGF was added to the bottom chamber as chemoattractants. After 24 h incubation, the migrated cells on the bottom surface were stained with 0.1% crystal violet (Santa Cruz Biotechnology, Inc.) and counted under an Olympus LCX100 Imaging system (Olympus Corporation, Tokyo, Japan).
Modified 24 well boyden chambers
Manufactured by Corning
Sourced in United States
The Modified 24-well Boyden chambers are a specialized lab equipment used to assess cell migration and invasion. The device consists of a two-chambered setup with a porous membrane separating the upper and lower compartments. Cells are seeded in the upper chamber, and the lower chamber contains a chemoattractant or other stimuli, allowing for the evaluation of cell movement through the membrane.
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2 protocols using modified 24 well boyden chambers
1
Cell Migration Assay with VEGF Chemoattractant
2
Endothelial Cell Migration Assay
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Cell migration assay was performed using modified 24-well Boyden chambers (Costar, New York, NY) containing a polycarbonate membrane with 5.0 μm pores. Mouse endothelial cells were starved in basic EBM2 (serum/growth factor-free) for 12 hours at 37°C before being harvested with trypsin and resuspended in EBM2 basal media with 5% of mouse plasma from the WT or the Cfh-/- mice. The single cell suspension was seeded at 1×105 cells/well in the upper chamber, and 0.5 ml EBM2 with 20 ng/ml of VEGF (Lonza) was added to the bottom chamber as chemoattractants. After 12 hrs, the migrated cells on the bottom surface were stained with 0.1% crystal violet and counted under microscope.
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