NK cells

The StraightFrom® CD56 MicroBead Kits were developed for the rapid selection and isolation of CD56⁺ cells directly from whole blood, buffy coat, or LRSC. The kits require no sample preparation and are optimized for a specific starting material. Subsequent depletion of CD3⁺ cells using CD3 MicroBeads, human yields purified NK cells.

The MACSxpress® NK Cell Isolation Kit,  human isolates untouched NK cells directly from small (2 mL) or larger (up to 30 mL) volumes of whole blood . Non-target cells are removed by immunomagnetic depletion. Simultaneously, erythrocytes are sedimented to yield untouched target cells of high purity.

Instead of working directly with whole blood or whole blood products, one can process the samples by density gradient centrifugation to pre-enrich peripheral blood mononuclear cells (PBMCs). PBMCs then serve as starting material for  NK and NKT cell isolation by the following options.

CD56 MicroBeads, human enable positive selection or depletion of CD56⁺ cells by direct magnetic labeling. Positive selection by the REAlease® CD56 MicroBead Kit, human, followed by depletion of CD3⁺ cells with CD3 MicroBeads, human leads to pure and non-activated NK cells.

The NK Cell Isolation Kit, human enables fast isolation of untouched NK cells from human PBMCs. Non-NK cells (i.e., T cells, monocytes, neutrophils, eosinophils, B cells, dendritic cells, granulocytes, and erythroid cells) are labeled with a cocktail of biotin-conjugated antibodies. Subsequently, these non-target cells are magnetically labeled with the NK Cell MicroBead Cocktail and depleted to yield highly pure NK cells.

The table summarizes dedicated kits for the effective isolation of different NK cell subsets. The new REAlease CD56 MicroBead Kit, human however enables the isolation of virtually any NK cell subset. The kit is designed to isolate CD56⁺ cells (NK and NKT cells) by positive selection. Magnetic labeling can then be easily removed for subsequent separation steps based on other markers.

Miltenyi Biotec offers a range of solutions for the analysis of NK cell–associated cytokines.

• MACS Antibodies can be used for intracellular staining of cytokines.
• MACS Cytokine Secretion Assays allow detection and enrichment of viable cytokine-secreting cells. The kits are available for a wide range of cytokines, including TNF-α, GM-CSF, IFN-α, IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-17, and IL-22, and can be used for further characterization of NK cell subsets.
• The MACSPlex Cytokine Kits are used for multiplex analysis of secreted cytokines in serum and cell culture supernatants using a standard flow cytometer. Cytokines that can be analyzed in a single sample include: GM-CSF, IFN-α, IFN-γ, lL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-17A, and TNF-α.

NK MACS Medium, research grade is a cell culture medium developed specifically for NK cells. It has been used in a variety of applications and, in combination with MACS Cytokines, is an ideal starting point for reliable cultivation conditions.

For detailed information about Miltenyi Biotec media optimized for NK cells, see chapter Cell culture media.

NK cell activation is essential for a variety of downstream application. Miltenyi Biotec offers polyclonal stimulation reagents that have been carefully designed to ensure optimal stimulation conditions.

The NK Cell Activation/Expansion Kit, human employs large cell-sized particles loaded with biotinylated antibodies against CD2 and CD335 (NKp46) to activate and expand primary cells. The large cell-sized particles mimic antigen-presenting cells and, when applied in a specific bead-to-cell ratio, lead to efficient NK cell activation.