Firefly luciferase-based chronological measurement of effector CD8 + T-cell activity using a multi-chamber luminometer.
Ryota HayashiHayato NakataniHinami KawahataRyonosuke FujieKaoru KurowarabeHaruko HayasakaPublished in: Bioanalysis (2023)
Background: Although cell-mediated cytotoxicity has been evaluated with various protocols, methods for monitoring cytotoxicity in a time series have not been established. This work describes a method for evaluating cytotoxicity using a multi-chamber real-time luminometer. Materials & methods: The efficiency of effector CD8 + T-cell expansion from melanoma-bearing splenocytes was analyzed. The effect of CD8 + T cells on the viability of luciferase-expressing target cells was measured by bioluminescence. Results: Melanoma-specific effector CD8 + T cells were differentiated by in vitro coculture. The melanoma cell growth was significantly inhibited in the presence of in vitro -expanded T cells in the bioluminescence-based time-lapse analysis. Conclusion: The bioluminescence-based assay is a useful method for monitoring the time course of cell viability of target tumor cells.
Keyphrases
- regulatory t cells
- dendritic cells
- energy transfer
- skin cancer
- type iii
- induced apoptosis
- single cell
- basal cell carcinoma
- high throughput
- cell cycle arrest
- stem cells
- signaling pathway
- cell proliferation
- immune response
- endoplasmic reticulum stress
- oxidative stress
- quantum dots
- mesenchymal stem cells
- bone marrow
- data analysis