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| Home > Publications database > Fura-10, unlike fura-2, is suitable for long-term calcium imaging in natural killer (NK) cells without compromising cytotoxicity and can be combined with target cell death analysis. |
| Home > Publications database > Fura-10, unlike fura-2, is suitable for long-term calcium imaging in natural killer (NK) cells without compromising cytotoxicity and can be combined with target cell death analysis. |
| Journal Article | DKFZ-2025-02655 |
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2025
Churchill Livingstone
Edinburgh [u.a.]
Abstract: There are compelling reasons to opt for primary human natural killer (NK) cells when validating Ca2+ indicators. 1.) NK cells exhibit a high degree of vulnerability to stressors such as indicator loading or light exposure. 2.) The lack of research on NK Ca2+ signaling underscores the necessity for developing reliable assays. 3.) The increased utilization of NK cell therapies necessitates a more profound comprehension of Ca2+ dependent signal transduction. Consequently, an assay was developed to monitor cytosolic Ca2+ signals in individual NK cells simultaneously with their cytotoxic function against cancer cells. We used this assay to assess the suitability of fura-2, fura-PE3, fura-8, fura-10 or fura-red for quantifying Ca2+ signals in NK cells without compromising their cytotoxic function. In contrast to the widely used fura-2, its red-shifted derivative fura-10 did not interfere with NK cytotoxicity over several hours. It exhibited a superior signal-to-noise ratio and good dynamic range, accompanied by minimal bleaching or leakage. Fura-8 and fura-red also preserved NK cell cytotoxicity, but had other disadvantages compared to fura-10. We successfully used fura-10 to report Ca2+ signals in NK cells from blood donors and patients diagnosed with lymphoma and leukemia over several hours at 37 °C during apoptotic or necrotic killing of different cancer cells (K562, THP1, OCI-AML2, and TMD8). Additionally, we show that fura-10 is well suited to report Ca2+ signals in intact murine pancreatic islets, another stress-sensitive cell preparation. Consequently, fura-10 is an optimal choice for measuring Ca²⁺ in primary human NK cells and other primary cell preparations.
Keyword(s): Calcium ; Cytotoxicity ; Fura-10 ; Fura-2 ; Natural killer (NK) cell ; Pancreatic islets ; cancer
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