JoVE Logo
Faculty Resource Center

Sign In

JoVE Encyclopedia of Experiments

Overview

Protocol

Encyclopedia of Experiments

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

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.

1. Calcein AM Staining-based Microscopic Method for Measuring NK Cell-mediated Cytotoxicity

  1. 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.
  2. 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.
  3. Add 1.5 µL of calcein AM solution (10 mM) to SK-HEP-1 ce.......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved