Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (4): 77-84.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0870

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Detection of NK Cell Cytotoxicity: Real-time Dynamic Imaging-Based Analysis

SANG Sen-hua()   

  1. Neukio(Shanghai)Biotherapeutics Co., Ltd., Shanghai 200000
  • Received:2023-09-11 Online:2024-04-26 Published:2024-04-30

Abstract:

Objective】Currently cellular immunotherapy is a fast-growing and reliable means of treating cancer. This study aims to develop a new method of detecting the effects of cell therapy products.【Method】By constructing tumor cells expressing green fluorescent protein(GFP), a method for evaluating the cytotoxicity of cell therapy products was constructed based on a real-time live-cell dynamic imaging system.【Result】The real-time live-cell dynamic imaging system was used to monitor K562-GFP cells, and there was a good linear relationship between the total fluorescence intensity and the number of cells. In co-culture with extremely low effector to target(E∶T)ratio, high accuracy was also achieved. The cytotoxicity of natural killer(NK)cells was accurately evaluated by calculating the half-maximum effector concentration(EC50)of NK cells for K562-GFP cells. Flow cytometry(FCM)was used to detect the correlation between apoptosis of K562-GFP cells and the quenching of cell fluorescent protein, which proved the feasibility of the detection method.【Conclusion】This method can be used to evaluate killing ability of immune cells of different types or production processes at a low cost, without requiring the use of dyes or antibodies.

Key words: cellular immunotherapy, apoptosis detection, NK cell, real-time dynamic imaging, half-maximum effector concentration