TY  - JOUR
TI  - Protocol for Isolation, Stimulation and Functional Profiling of Primary and iPSC-derived Human NK Cells
AU  - Melsen, Janine 
AU  - Themeli, Maria 
AU  - van Ostaijen-ten Dam, Monique  
AU  - van Beelen, Els 
AU  - Lugthart, Gertjan 
AU  - Hoeben, Rob 
AU  - Schilham, Marco 
AU  - Mikkers, Harald  
VL  - 10
IS  - 23
PY  - 2020
DA  - 2020/12/05
SP  - e3845
C1  - Bio-protocol 2020;10:e3845
DO  - 10.21769/BioProtoc.3845
UR  - https://doi.org/10.21769/BioProtoc.3845
AB  - Natural killer (NK) cells are innate immune cells, characterized by their cytotoxic capacity, and chemokine and cytokine secretion upon activation. Human NK cells are identified by CD56 expression. Circulating NK cells can be further subdivided into the CD56bright (~10%) and CD56dim NK cell subsets (~90%). NK cell-like cells can also be derived from human induced pluripotent stem cells (iPSC). To study the chemokine and cytokine secretion profile of the distinct heterogenous NK cell subsets, intracellular flow cytometry staining can be performed. However, this assay is challenging when the starting material is limited. Alternatively, NK cell subsets can be enriched, sorted, stimulated, and functionally profiled by measuring secreted effector molecules in the supernatant by Luminex. Here, we provide a rapid and straightforward protocol for the isolation and stimulation of primary NK cells or iPSC-derived NK cell-like cells, and subsequent detection of secreted cytokines and chemokines, which is also applicable for a low number of cells.
KW  - CD56
KW  - Natural killer cells
KW  - Induced pluripotent stem cells
KW  - Cytokines
KW  - Chemokines
KW  - Peripheral blood
KW  - Luminex
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False