Measurement of Natural Killer Cell Migration in Response to Chemotactic Stimuli
Published: November 30, 2023
Abstract
Source: Chava, S., et al. Measurement of Natural Killer Cell-Mediated Cytotoxicity and Migration in the Context of Hepatic Tumor Cells. 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.
Protocol
1. NK Cell Migration Assay Grow NK92MI cells and centrifuge cells at 160 x g for 3 min in a 15 mL sterile centrifuge tube. 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. Plate NK92MI cells (2.5 x 105 cells/100 µL/well) in the upper compartment of the transwell permeable chamber (6.5 mm diameter insert and 5 µ…
Divulgaciones
The authors have nothing to disclose.
Materials
| Absolute counting beads | Thermo Fisher Scientific | C36950 | |
| NK92MI cells | ATCC | CRL-2408 | |
| SKHEP-1 Cells | ATCC | HTB-52 | |
| Transwell permeable chambers | Costar | 3241 | |
| Opti-MEM | Gibco | 31985070 | |
| Phosphate Buffer Saline (PBS) | Sigma-Aldrich | P4417 | |
| Alpha-MEM | Sigma-Aldrich | M4256 |
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Measurement of Natural Killer Cell Migration in Response to Chemotactic Stimuli. J. Vis. Exp. (Pending Publication), e21784, doi: (2023).