%0 Journal Article
%A Kaschek, Lea
%A Vialle, Joanne
%A Stopper, Gebhard
%A Hoffmann, Markus D A
%A Zöphel, Sylvia
%A Jansky, Johanna
%A Küchler, Nadja
%A Schäfer, Gertrud
%A Moter, Alina
%A Wendel, Philipp
%A Neumann, Frank
%A Schormann, Claudia
%A Ullrich, Evelyn
%A Roma, Leticia Prates
%A Kummerow, Carsten
%A Thurner, Lorenz
%A Schwarz, Eva C
%A Hoth, Markus
%T 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.
%J Cell calcium
%V 133
%@ 0143-4160
%C Edinburgh [u.a.]
%I Churchill Livingstone
%M DKFZ-2025-02655
%P 103091
%D 2025
%X 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.
%K Calcium (Other)
%K Cytotoxicity (Other)
%K Fura-10 (Other)
%K Fura-2 (Other)
%K Natural killer (NK) cell (Other)
%K Pancreatic islets (Other)
%K cancer (Other)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:41314008
%R 10.1016/j.ceca.2025.103091
%U https://inrepo02.dkfz.de/record/306662