A human leukemic cell line, THP-1 cell (American Type Culture Collection, Manassas, VA, USA), was cultured in RPMI 1640 medium containing 10% fetal bovine serum, 20 µg/mL penicillin and 20 µg/mL streptomycin at 37°C in a humidified atmosphere with 5% CO2. The cells were differentiated into macrophages by adding 50 ng/mL of phorbol-12-myristate-13-acetate (PMA; EMD Biosciences, La Jolla, CA, USA) for 48 hours in 35 mm Petri dishes. After differentiation, THP-1 cells were, respectively, treated with 10, 20, 50 and 100 ng/mL of recombinant human IL-17 (R&D Systems, Inc., Min-neapolis, MN, USA) for 48 hours. Then, the THP-1 cells were treated with 100 ng/mL of recombinant human IL-17 for 0, 24 and 48 hours, respectively. To inhibit NF-κB pathways, the cells were incubated with 10 mM BAY11-7082 (Beyotime Institute of Biotechnology, Jiangsu, China) accompanied by IL-17 for 48 hours.
Thp 1 cells
Manufactured by R&D Systems
Sourced in United States
THP-1 cells are a human monocytic cell line derived from the peripheral blood of a 1-year-old male with acute monocytic leukemia. They can be used as a model for studying monocyte and macrophage biology, as well as inflammatory responses.
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Lab products found in correlation
Thp 1 cell, by American Type Culture Collection (1 mentions)
Recombinant human il 17, by R&D Systems (1 mentions)
Il 17, by Beyotime (1 mentions)
Bay 11 7082, by Beyotime (1 mentions)
Facscanto 2, by BD (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Facsdiva software, by BD (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
96 well plate, by Corning (1 mentions)
Thp 1 cell, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Af2148, by R&D Systems (1 mentions)
Fetal calf serum (fcs), by Biosera (1 mentions)
2 protocols using thp 1 cells
1
Differentiation of THP-1 Cells into Macrophages Treated with IL-17
2
THP1 Lipoprotein-αGalCer Uptake Assay
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THP1 cells (American Type Culture Collection), known for their constitutive LDLR expression (17 (link)), were cultured in RPMI 1640 (Gibco, Thermo Fisher Scientific) supplemented with 10% heat-inactivated FCS (ultra-low endotoxin, Biosera), 4 mmol/L Glutamax (Gibco, Thermo Fisher Scientific), 5 mmol/L sodium pyruvate (MilliporeSigma), and antibiotics (Gibco, Thermo Fisher Scientific). For the uptake experiments, THP1 cells were washed in PBS, and 100,000 THP1 cells were seeded in 100 μL Opti-MEM (Gibco, Thermo Fisher Scientific) on a 96-well plate (Corning). Upon preincubation with 5 μg/mL LDLR-blocking antibody for 30 minutes (AF2148, R&D Systems, Bio-Techne), THP1 cells were incubated for 4 hours with 0.3 μL (6 μg/mL) or 0.03 μL (0.6 μg/mL) lipoprotein-αGalCer complexes per well, respectively, containing 100 ng and 10 ng αGalCer. Finally, loading of the DyLight 633–lipoprotein and AF488-αGalCer was assessed on a FACSCanto II flow cytometer (BD Biosciences) with FACSDiva software (BD Biosciences), and data were analyzed with FlowJo software v10.
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