Start by adding chemoattractants, such as a chemokine, in a multiwell plate.
Place a transwell permeable chamber of an appropriate pore size into the well. Add natural killer, or NK, cell suspension to the upper chamber and incubate.
A few chemokine molecules diffuse through the membrane and bind to NK cell receptors. This triggers signaling pathways that promote cytoskeletal changes, resulting in directional cell movement toward the higher chemoattractant concentration in the lower chamber.
After incubation, transfer a fixed volume of migrated cells from the lower chamber to a fluorescence-activated cell sorting or FACS tube. Add a predetermined number of fluorescent counting beads and perform FACS-based cell counting.
Using the following formula, determine the absolute number of migrated NK cells in the sample.
The dot plot gives the number of counting beads and cells separated as distinct regions, based on their fluorescence and scatter properties.
Evaluation of the Cell Invasion and Migration Process: A Comparison of the Video Microscope-based Scratch Wound Assay and the Boyden Chamber Assay
Live-Cell Imaging Assays to Study Glioblastoma Brain Tumor Stem Cell Migration and Invasion
A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion
Copyright © 2024 MyJoVE Corporation. All rights reserved