Protocol for Isolation, Stimulation and Functional Profiling of Primary and iPSC-derived Human NK Cells

BIO-PROTOCOL, vol. 10, pp. e3845

Abstract

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.

Authors 8

1. Janine E. Melsen Aachen Department of Pediatrics, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands;

2. Maria Themeli

   Amsterdam University Medical Centers

   Affiliation as printed

   Department of Hematology, Amsterdam UMC, Amsterdam, The Netherlands

   Department of Hematology, Amsterdam UMC, Amsterdam, The Netherlands;

3. M. van Dam Aachen Department of Pediatrics, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands;

4. Els van Beelen Aachen Department of Immunohematology and Blood Transfusion

   Leiden University Medical Center

   Affiliation as printed

   Department of Immunohematology & Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands

   Department of Immunohematology & Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands;

5. Gertjan Lugthart Aachen Department of Pediatrics, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands;

6. Rob C. Hoeben Aachen Department of Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Department of Cell & Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands

7. Marco W. Schilham Aachen Department of Pediatrics, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands

   Department of Pediatrics, Leiden University Medical Center, Leiden, The Netherlands;

8. Harald M. M. Mikkers Aachen Department of Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Department of Cell & Chemical Biology, Leiden University Medical Center, Leiden, The Netherlands