X-VIVOTM 10 Serum-free Hematopoietic Cell Medium

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	supplemented with 450 U/ml of IL-2 (Proleukin; Novartis),

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supplemented with 450 U/ml of IL-2 (Proleukin; Novartis),

Publication protocol

The HL cell lines KM-H2, L-428, and L-540, the multiple myeloma cell lines RPMI-8226 and U266, and the AML cell line K562 were cultured in RPMI-1640 (Sigma-Aldrich; St. Louis, MO) supplemented with 10% fetal bovine serum (Gibco; Gaitherburg, MD) at 37 °C in a humidified atmosphere with 5% CO2. The NK cell lines, NK-92 and haNK (kindly provided by NantKwest, Inc.), were cultured in X-VIVO 10 (Lonza; Anaheim, CA) plus 5% heat-inactivated human AB serum at 37 °C in a humidified atmosphere with 5% CO2. NK-92 cell culture media was supplemented with 450 U/ml of IL-2 (Proleukin; Novartis), while haNK cells were cultured without IL-2 because of self-production capacity. Primary NK cells were obtained from two different healthy donors. Peripheral blood mononuclear cells were isolated by Lymphoprep (Stemcell Technologies; Vancouver, BC) density gradient centrifugation. Subsequently, NK cells were negatively enriched using a NK Cell Isolation Kit (Stemcell Technologies) according to the manufacturer’s instruction. Isolated NK cells were incubated in AIM-V medium supplemented with 10% human AB serum (Sigma-Aldrich) and 10 ng/mL recombinant human IL-15 (Peprotech; Rocky Hill, NJ) for 2 weeks. Half volume of the medium was replaced twice a week. All experiments using cell lines were performed within 12 weeks of cell culture.

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Download the product protocol from Lonza for X-VIVOTM 10 Serum-free Hematopoietic Cell Medium below.

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