Human PBMCs from healthy volunteers were isolated from heparinized venous blood using Ficoll-Hypaque (Lymphoprep; Alere technologies, Oslo, Norway) as described previously.18 (link) For monocyte-derived macrophages (MDMs) differentiation, adherent monocytes were incubated in Roswell Park Memorial Institute 1640 medium (Lonza, Basel, Switzerland) containing 5% pooled human serum (Sigma-Aldrich, St. Louis, MO, USA), 1% L-glutamine, for 1 hour at 37°C, after which the nonadherent cells were removed. Human MDMs were prepared by culturing peripheral blood monocytes for 4 days in the presence of 4 ng/mL human macrophage colony-stimulating factor (R&D Systems, Minneapolis, MN, USA). SARS-CoV-2 (2019-nCoV) NC-His recombinant protein (cat. No. 40588-V08B), Spike S1-His recombinant protein (cat. No. 40591-V08H), Spike S2 extracellular domain (ECD)-His recombinant protein (cat. No. 40590-V08B), and Spike RBD-His recombinant protein (cat. No. 40592-V08H) were purchased from Sino Biological, Beijing, China. Cells were stimulated with the proteins as indicated in figure legends.
Ficoll hypaque lymphoprep
Manufactured by Abbott
Sourced in Norway
Ficoll-Hypaque (Lymphoprep) is a density gradient medium used for the isolation of mononuclear cells, such as lymphocytes and monocytes, from whole blood or other cell suspensions. It is a sterile, pyrogen-free solution composed of Ficoll and sodium diatrizoate.
Automatically generated - may contain errors
2 protocols using ficoll hypaque lymphoprep
1
Isolation and Differentiation of Human Monocyte-derived Macrophages
2
PBMC and Decidua Isolation
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Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll Hypaque (Lymphoprep™; Alere Technologies, Norway) density gradient centrifugation. First trimester decidua was isolated from uterine content that was collected by induced abortion. Third trimester decidua was dissected from maternal surface of the delivered placenta. Decidua was rinsed with phosphate buffered saline (PBS) until the blood was removed, minced with a pair of scissors to produce 1–2 mm pieces, and filtered through 32-μm nylon mesh. All samples were cryopreserved.
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