Isolation of natural killer (NK) cells from human blood products
Philippa R Kennedy
Abstract
Standard isolation procedure for peripheral blood mononuclear cells from human blood leukopheresis products
Steps
Healthy donor blood products (Leukopaks) were obtained from Memorial Blood Bank (Minneapolis, MN). All samples were de-identified and their use was approved by the University of Minnesota and NMDP institutional review board in accordance with the Declaration of Helsinki.
Peripheral blood mononuclear cells (PBMCs) were separated through density gradient filtratation (Ficoll-Paque; GE Healthcare).
For experiments with enriched NK cells, PBMCs were processed fresh using negative magnetic bead separation (EasySep Human NK Cell Enrichment Kit; Cat. No:19055, STEMCELL Technologies).
Experiments with PBMCs or enriched NK cells were performed in RPMI 1640 supplemented with 10% Fetal Bovine Serum and 1% penicillin/streptomycin. Where specified, culture was supplemented with recombinant human IL-15 (National Cancer Institute).
