Isolation and Purification of Human NK Cells

Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by centrifugation over a density gradient medium (Ficoll-Paque Premium; GE Healthcare, Uppsala, Sweden) to separate granulocytes as previously described [27 (link), 28 (link)]. PBMCs were stained with trypan blue stain (Invitrogen, Carlsbad, CA) to determine total cell count and cell viability and adjusted to a final concentration of 5 × 10⁷ cells/ml. NK cells were isolated from PBMCs using an EasySep Negative Human NK Cell Isolation Kit (Stemcell Technologies, Vancouver, BC, Canada) as previously described [27 (link), 28 (link)].

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Balinas C., Cabanas H., Staines D, & Marshall-Gradisnik S. (2019). Identification and characterisation of transient receptor potential melastatin 2 and CD38 channels on natural killer cells using the novel application of flow cytometry. BMC Immunology, 20, 14.

Publication 2019

Ficoll-Paque Premium trypan blue stain EasySep Negative Human NK Cell Isolation Kit

Blood Cell isolation Cell viability Cells Density gradient centrifugation Ficoll Granulocytes Human Nk cells Peripheral blood mononuclear cells Stain Stemcell Trypan blue

Corresponding Organization : Griffith University

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Variable analysis

independent variables

Isolation of peripheral blood mononuclear cells (PBMCs) from whole blood using density gradient centrifugation
Isolation of natural killer (NK) cells from PBMCs using negative selection kit

dependent variables

Total cell count and cell viability of PBMCs

control variables

Ficoll-Paque Premium density gradient medium used for PBMC isolation
EasySep Negative Human NK Cell Isolation Kit used for NK cell isolation
Final concentration of PBMCs adjusted to 5 × 10^7 cells/ml

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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