Measurement of Natural Killer Cell Migration in Response to Chemotactic Stimuli

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

1. Grow NK92MI cells and centrifuge cells at 160 x g for 3 min in a 15 mL sterile centrifuge tube.
2. 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.
3. 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 µm pore size).
4. In the bottom chamber add 0.6 mL of serum-free medium containing material to be tested for NK cell chemoattractant properties ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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