BMDMs were obtained using L929 cell-conditioned medium, as previously described46 (link). Briefly, bone marrow cells were obtained by flushing both mice femurs and tibias with RPMI-1640 culture media (Corning, 15-040-CV). After centrifugation, cells were resuspended in RPMI-1640 containing 20% L929, 10% foetal bovine serum (FBS, GE Life Sciences, SV30160.03), L-glutamine (2 mM, Sigma, G7513), penicillin (100 U/mL, Sigma, P4333), and fungizone (2.5 μg/mL, Gibco, 15-290-018) and seeded for 7 days. After the end of this period, BMDMs were harvested for subsequent experiments.
Rpmi 1640 culture media
Manufactured by Corning
Sourced in United States
RPMI-1640 is a cell culture medium that provides essential nutrients to support the growth and maintenance of a variety of cell types, including those derived from human tissues. It is a widely used basal medium in cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Fungizone, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
P4333, by Thermo Fisher Scientific (1 mentions)
Human ifn γ elisa kit, by MultiSciences Biotech (1 mentions)
24 well plate, by Corning (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
2 protocols using rpmi 1640 culture media
1
Differentiation of Bone Marrow-Derived Macrophages
2
Blocking IFN-γ Induction by N-IgY
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The project of this in vitro experiment was approved by the ethics committee of West China Hospital of Stomatology, Sichuan University (WCHSIRB-D-2020–355). Samples of anticoagulated peripheral blood were donated by healthy human donors (n = 7). Basic characteristics of the donors were listed in Tab. S1. Peripheral blood mononuclear cells (PBMCs) were separated by Ficoll reagent (Haoyang, China) under density gradient centrifugation according to the operating manual. The induction of interferon-γ (IFN-γ) was then measured to identify the blocking ability of N-IgY.
The isolated PBMCs were incubated in RPMI 1640 culture media (Corning, USA) supplemented with 10% of FBS (Corning, USA). The cells were seeded (5 × 105 cells/500 μL/well) in 24-well plate (Corning, USA) and cultured at 37 °C in a 5% humidified CO2 atmosphere.
Cells were treated with NP (20 μg/mL) alone (groupN ), or NP premixed with N-IgY (1 mg/mL) for 30 min (group N + IgY ) for 18 h. Those treated with PBS were set as control (group Con ). After centrifugation at 4000 rpm and 4 °C for 3 min, the separated supernatant was immediately frozen at −20 °C until detected. Human IFN-γ ELISA kit (MULTI SCIENCES, China) was used to detect the secretion of IFN-γ in the culture supernatant according to the operating manual.
The isolated PBMCs were incubated in RPMI 1640 culture media (Corning, USA) supplemented with 10% of FBS (Corning, USA). The cells were seeded (5 × 105 cells/500 μL/well) in 24-well plate (Corning, USA) and cultured at 37 °C in a 5% humidified CO2 atmosphere.
Cells were treated with NP (20 μg/mL) alone (group
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