Male C57BL/6NCr mice were purchased from the National Cancer Institute (NCI) Animal Production Program and Charles River Laboratories International (Frederick, MD) and used between 24–25 weeks of age. Male C57BL/6-Tg(UBC-GFP)30Scha/J mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and used between the 6–12 weeks of age. Mice were stored in a pathogen-free facility throughout the entirety of the study.
NK cells were isolated by harvesting the spleen and bone marrow and processing them into single-cell suspensions. Erythrocytes were depleted using ACK lysing solution (Lonza, Walkersville, MD). NK cells were isolated by negative selection using magnetic cell separation beads (Miltenyi Biotec, cat#: 130-115-818), sorted using AutoMACS® Separator (Miltenyi Biotec, San Diego, CA), and cultured at 37°C in 5% CO2 in complete mouse media33 (link) (CMM) supplemented with 100 IU/mL recombinant human IL-2 (NCI Biological Resources Branch, Frederick, MD).
NK cells were isolated by harvesting the spleen and bone marrow and processing them into single-cell suspensions. Erythrocytes were depleted using ACK lysing solution (Lonza, Walkersville, MD). NK cells were isolated by negative selection using magnetic cell separation beads (Miltenyi Biotec, cat#: 130-115-818), sorted using AutoMACS® Separator (Miltenyi Biotec, San Diego, CA), and cultured at 37°C in 5% CO2 in complete mouse media33 (link) (CMM) supplemented with 100 IU/mL recombinant human IL-2 (NCI Biological Resources Branch, Frederick, MD).