Author: Ma, Ge; Greenwell-Wild, Teresa; Lei, Kejian; Jin, Wenwen; Swisher, Jennifer; Hardegen, Neil; Wild, Carl T.; Wahl, Sharon M.
Title: Secretory Leukocyte Protease Inhibitor Binds to Annexin II, a Cofactor for Macrophage HIV-1 Infection Document date: 2004_11_15
ID: rlabxfss_4
Snippet: Isolation of Monocytes. Human PBMCs, obtained by leukapheresis of healthy volunteers (Dept. of Transfusion Medicine, NIH), were diluted in endotoxin-free PBS without Ca 2 Ï© and Mg 2 Ï© for density sedimentation. Monocytes and T cells were purified by elutriation (2, 3) . Monocytes were cultured in DMEM (BioWhittaker) supplemented with 2 mM l -glutamine and 10 g/ml gentamicin. For monocyte-derived macrophages, monocytes were adhered and cultured .....
Document: Isolation of Monocytes. Human PBMCs, obtained by leukapheresis of healthy volunteers (Dept. of Transfusion Medicine, NIH), were diluted in endotoxin-free PBS without Ca 2 ϩ and Mg 2 ϩ for density sedimentation. Monocytes and T cells were purified by elutriation (2, 3) . Monocytes were cultured in DMEM (BioWhittaker) supplemented with 2 mM l -glutamine and 10 g/ml gentamicin. For monocyte-derived macrophages, monocytes were adhered and cultured in supplemented DMEM and 10% FBS for 7-10 d. The human myelomonocytic U937 cell line (American Type Culture Collection) was cultured in RPMI 1640 medium containing 2 mM l -glutamine, 50 IU/ml penicillin, 50 g/ml streptomycin, and 10% FBS at 37 Њ C in a humidified atmosphere of 5% CO 2 .
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