Isolation, expansion and flow analysis of NK cells in COVID-19 research 

Isolation, expansion and flow analysis of NK cells in COVID-19 research 

NK cells play a primary role as a first line of defence by mediating anti-viral immune responses. Isolation and expansion of NK cells from virus-infected individuals are valuable steps for studying their immune responses i.e. by flow cytometry.  

NK Cell Isolation 

The MACSxpress® Whole Blood NK Cell Isolation Kit, human enables fast isolation of NK cells with high purity from as little as 2 mL of whole blood. MACSxpress Technology is designed to minimize hands-on time – density-gradient centrifugation and erythrocyte lysis are not required. 

Before separation
After separation

Untouched NK cells from whole blood. The MACSxpress  Whole  Blood NK Cell Isolation Kit, human, was used to separate NK cells from 30 mL of human EDTA-anticoagulated whole blood. The isolated cells were fluorescently stained with CD45-VioBlue, CD3-FITC, and CD56-PE, and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, non-leukocytes, and dead cells were excluded from the analysis based on CD45 expression, scatter signals, and propidium iodide fluorescence.

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         Fast isolation of NK cells from human whole blood

NK cell cultivation

Comprehensive assays require a large quantity of NK cells. NK MACS® Medium has been optimized specifically for NK cell cultivation and expansion. Cells expanded in this medium remain fully functional and are ready to use for any downstream assay.

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.

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Superior expansion of NK cells in NK MACS Medium. NK cell expansion rate from (A) PBMCs (n = 3) or (B) isolated NK cells (n = 3) using 5% AB serum and 500 IU/mL of IL-2 is significantly higher in NK MACS Medium compared to standard culture methods. 

Flow analysis of NK cells

  • NK cells play an important role against COVID-19, latest research shows function of NK and CD8+ T cells  was exhausted with the increased expression of NKG2A (CD159a) in COVID-19 patients. (Zheng et al. PMID: 32203188) and Tim-3 (CD366) (Maucaurant et al. PMID: 32826343).
  • MACSplex Assays can be used if you are interested in analysis of multiple soluble cytokines in one sample.
  • The 8-Color Immunophenotyping Kit, anti-human, REAfinity is a pre-titrated antibody cocktail which offers the most reliable immunophenotyping in a variety of samples. This enables you to identify human immune cell subsets such as monocytes, neutrophils, eosinophils, T cells, B cells, and NK cells in one sample – quickly, easily, and reproducibly.
  • Download the Application Note, allowing comprehensive phenotypic analysis of NK cells

Check out more supporting materials for your NK cell research

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