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Cell staining

NS Nikita Subedi
LE Laura C. Van Eyndhoven
AH Ayla M. Hokke
LH Lars Houben
MT Mark C. Van Turnhout
CB Carlijn V. C. Bouten
KE Klaus Eyer
JT Jurjen Tel
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5 µM Calcein Red AM (AAT Bioquest, Catalog no. 21900) and 10 µM Cell Tracker Blue dyes (Invitrogen, C2110) were used to stain primary NK cells and target cells (K562 or Jurkat), respectively. For staining, around 2 million cells were washed with RPMI free of supplements and Phenol red (Gibco, Catalog no. 11835030), resuspended in the freshly prepared dye solution in-RPMI (1 mL) and incubated at 37 °C for 30 min. Thereafter, cells were subsequently washed twice and resuspended in RPMI medium supplemented with 25 mM HEPES (Gibco, Catalog no. 15630056), 2% human serum (Sanquin), without Phenol red) containing 2.5 µM Sytox green (Invitrogen, Catalog no. S7020) and 7 µM CellEvent Caspase-3/7 Green Detection Reagent (Invitrogen, Catalog no. C10423), respectively. The stained cells were then proceeded for cell encapsulation.

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