TY  - JOUR
AU  - Kaschek, Lea
AU  - Vialle, Joanne
AU  - Stopper, Gebhard
AU  - Hoffmann, Markus D A
AU  - Zöphel, Sylvia
AU  - Jansky, Johanna
AU  - Küchler, Nadja
AU  - Schäfer, Gertrud
AU  - Moter, Alina
AU  - Wendel, Philipp
AU  - Neumann, Frank
AU  - Schormann, Claudia
AU  - Ullrich, Evelyn
AU  - Roma, Leticia Prates
AU  - Kummerow, Carsten
AU  - Thurner, Lorenz
AU  - Schwarz, Eva C
AU  - Hoth, Markus
TI  - 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.
JO  - Cell calcium
VL  - 133
SN  - 0143-4160
CY  - Edinburgh [u.a.]
PB  - Churchill Livingstone
M1  - DKFZ-2025-02655
SP  - 103091
PY  - 2025
AB  - 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.
KW  - Calcium (Other)
KW  - Cytotoxicity (Other)
KW  - Fura-10 (Other)
KW  - Fura-2 (Other)
KW  - Natural killer (NK) cell (Other)
KW  - Pancreatic islets (Other)
KW  - cancer (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:41314008
DO  - DOI:10.1016/j.ceca.2025.103091
UR  - https://inrepo02.dkfz.de/record/306662
ER  -