eoe sub articles

EoE Sub Articles

1. NK Cell Migration Assay
<ol>
	<li>Grow NK92MI cells and centrifuge cells at 160 x g for 3 min in a 15 mL sterile centrifuge tube.</li>
	<li>Wash the cell pellet twice with 5 mL of 1x PBS and resuspend the cells in 3 mL of serum-free NK92MI cell medium. Count NK92MI cells using a hemocytometer or an automated cell counter.</li>
	<li>Plate NK92MI cells (2.5 x 105 cells/100 &#181;L/well) in the upper compartment of the transwell permeable chamber (6.5 mm diameter insert and 5 &#181;...

measurement of natural killer cell migration in response to chemotactic stimuli

Source: Chava, S., et al. <a href="https://app.jove.com/v/60714">Measurement of Natural Killer Cell-Mediated Cytotoxicity and Migration in the Context of Hepatic Tumor Cells.</a> J. Vis. Exp. (2020).
This video demonstrates a method to study natural killer (NK) cell migration through a porous membrane insert system in response to chemoattractants. This method enables quantitative analysis of NK cell migration using flow cytometry.

Measurement of Natural Killer Cell Migration in Response to Chemotactic Stimuli

Source: Chava, S., et al. Measurement of Natural Killer Cell-Mediated Cytotoxicity and Migration in the Context of Hepatic Tumor Cells. J. Vis. Exp. ...

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.