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Firefly luciferase-based chronological measurement of effector CD8 + T-cell activity using a multi-chamber luminometer.

Ryota HayashiHayato NakataniHinami KawahataRyonosuke FujieKaoru KurowarabeHaruko Hayasaka
Published 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
  • high throughput
  • cell cycle arrest
  • basal cell carcinoma
  • stem cells
  • oxidative stress
  • mesenchymal stem cells
  • bone marrow