Calcein Acetoxymethyl Staining to Evaluate Natural Killer Cell-Mediated Cytotoxicity toward Cancer Cells

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 an assay to measure the cytotoxicity of natural killer cells against cancer cells using the Calcein AM stain. The reduced number of fluorescent cancer cells following co-culture with natural killer cells indicates natural killer cell-mediated cytotoxicity toward cancer cells.

Protocol

1. Calcein AM Staining-based Microscopic Method for Measuring NK Cell-mediated Cytotoxicity Culture SK-HEP-1 cells to 70−80% confluency. Generate a single-cell suspension by first washing cells with 5 mL of 1x PBS followed by incubation with 1 mL of 0.25% trypsin-EDTA. Centrifuge SK-HEP-1 cells in a 1 mL sterile centrifuge tube at 160 x g for 3 min. Resuspend the pellet in 3 mL of serum-free DMEM. Add 1.5 µL of calcein AM solution (10 mM) to SK-HEP-1 ce…

Divulgaciones

The authors have nothing to disclose.

Materials

Calcein AM dye Sigma-Aldrich 17783
DMEM Gibco 11965-092
Fetal Bovine Serum Gibco 26140079
Horse Serum Gibco 16050114
NK92MI cells ATCC CRL-2408
Phosphate Buffer Saline (PBS) Sigma-Aldrich P4417
SKHEP-1 Cells ATCC HTB-52
Trypsin EDTA solution Gibco 25200056

Tags

Play Video

Citar este artículo
Calcein Acetoxymethyl Staining to Evaluate Natural Killer Cell-Mediated Cytotoxicity toward Cancer Cells. J. Vis. Exp. (Pending Publication), e21783, doi: (2023).

View Video