An In Vitro Assay for Evaluating Natural Killer Cell Cytotoxicity against Cancer Cells
Published: May 31, 2024
Abstract
Source: Baik, C. H., et al. A Plate-based Cytotoxicity Assay for the Assessment of Rat Placental Natural Killer Cell Cytolytic Function. J. Vis. Exp. (2019)
This video demonstrates performing a lactate dehydrogenase release assay to determine the cytotoxicity of natural killer (NK) cells against the target cancer cells. The assay quantifies the release of the cytoplasmic lactate dehydrogenase enzyme from cancer cells by measuring the absorbance of the red-colored product formed.
Protocol
1. Cytotoxicity Assay: Assay Protocol Use a round bottom, culture treated 96-well plate to set up the plate as suggested in Table 1. This table shows the experimental controls and 3 sets of NK experimental columns. This can be expanded to a total of 10 NK experimental columns in a 96-well plate. Centrifuge the assay plate at 250 x g for 4 min to be certain that the effector and target cells are in contact. Incubate the 96-well plate for 5 hours in a humidified chamb…
Divulgations
The authors have nothing to disclose.
Materials
| 96-well Tissue Culture Plate | CELLTREAT | 229190 | Sterile, Round Bottom |
| Cell scraper | Fisher | 8100241 | |
| Cytotox 96 Non-Radioactive Cytotoxicity Assay Kit | Promega | G1780 | |
| RPMI | Gibco | 11875135 | 1640 Medium |
| YAC-1 cell | ATCC | TIB-160 |
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Citer Cet Article
An In Vitro Assay for Evaluating Natural Killer Cell Cytotoxicity against Cancer Cells. J. Vis. Exp. (Pending Publication), e22259, doi: (2024).