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).

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