Isolation and Preservation of NK Cells

Peripheral blood mononuclear cells (PBMCs) were isolated using a Lymphoprep^™ (Gibco) centrifugation gradient and half of the PBMCs were used directly for NK cell enrichment as previously described [21 (link),22 (link)]. NK cells were kept at a concentration of 10⁶ NK cells/ml in media (RPMI-1640 with L-Glutamine (Gibco) with 10% heat-inactivated fetal bovine serum (FBS; Gibco) and 1% Penicillin/Streptomycin (Gibco)) until analysis. The other half of the PBMCs was frozen based on a method by Paich et. al. [23 (link)]).

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Müller L., Meyer M., Bauer R.N., Zhou H., Zhang H., Jones S., Robinette C., Noah T.L, & Jaspers I. (2016). Effect of Broccoli Sprouts and Live Attenuated Influenza Virus on Peripheral Blood Natural Killer Cells: A Randomized, Double-Blind Study. PLoS ONE, 11(1), e0147742.

Publication 2016

Lymphoprep™ RPMI-1640 L-Glutamine fetal bovine serum (FBS; Penicillin/Streptomycin

Centrifugation Frozen Glutamine Lymphoprep Nk cells Penicillin Peripheral blood mononuclear cells Streptomycin

Corresponding Organization : Stanford University

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Variable analysis

independent variables

Isolation of peripheral blood mononuclear cells (PBMCs) using Lymphoprep™ gradient centrifugation
Enrichment of natural killer (NK) cells from PBMCs

dependent variables

NK cell concentration (10^6 NK cells/ml)

control variables

NK cell media (RPMI-1640 with L-Glutamine, 10% heat-inactivated fetal bovine serum, 1% Penicillin/Streptomycin)
Freezing of the other half of PBMCs

positive controls

None specified

negative controls

None specified

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