Human THP-1 monocytic leukemia cells were purchased from American Type Culture Collection (Manassas, VA, USA) and labeled with 3 μM BCECF-AM (Dojindo, Kumamoto, Japan) as described previously [21 (link)]. HUVECs were treated with 100 μg/ml AGE-BSA or non-glycated BSA in the presence or absence of 670 ng/ml n-butanol extracts of noni for 24 h and then incubated with BCECF-AM-labeled THP-1 cells for 4 h. After the incubation, non-adherent THP-1 cells were removed, and fluorescent intensities of the adherent THP-1 cells were measured as described previously [21 (link)].
Human thp 1 monocytic leukemia cells
Manufactured by American Type Culture Collection
Sourced in United States
The Human THP-1 monocytic leukemia cells are a well-established in vitro model derived from a human acute monocytic leukemia patient. These cells possess characteristics of monocytes and are commonly used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Bcecf am, by Dojindo Laboratories (2 mentions)
Fetal bovine serum (fbs), by Nichirei Biosciences (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Heat inactivated fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
5 protocols using human thp 1 monocytic leukemia cells
1
Quantifying AGE-Induced Monocyte Adhesion
2
THP-1 Monocyte Adhesion Assay
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Human THP-1 monocytic leukemia cells (American Type Culture Collection, Manassas, VA, USA) were maintained in RPMI 1640 medium supplemented with 1% GultaMAX (Life Technologies Corporation, Carlsbad, CA, USA) and 1% fetal bovine serum (NICHIREI BIOSCIENCES INC, Tokyo, Japan). THP-1 cells were labeled with 3 mM BCECF-AM (Dojindo, Kumamoto, Japan) at 37°C for 30 min according to the supplier’s recommendation. THP-1 cell adhesion to HUVECs was assayed according to the method described previously [17 (link)]. Briefly, HUVECs were treated with or without 100 μg/ml non-glycated BSA or 3% citrated human plasma in the presence or absence of 30 nM rivaroxaban for 4 hr, and then incubated with BCECF-AM-labeled THP-1 cells for 4 hr. After the incubation, nonadherent THP-1 cells were removed by washing the HUVECs gently. Fluorescent intensities of the adherent THP-1 cells were measured.
3
Monocyte and Macrophage Differentiation Assay
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Human THP-1 monocytic leukemia cells were obtained from the American Type Culture Collection. THP-1 cells were maintained in RPMI 1640 medium with 10% heat-inactivated FCS, 50 U/mL of penicillin G, and 50 mg/mL of streptomycin sulfate in an atmosphere containing 5% CO2 and 95% air. Cultures were maintained at a cell concentration between 2 × 105 and 1 × 106 cells/mL, with culture medium added every 2 days. Treating THP-1 cells with 200 nM phorbol myristate acetate (PMA) for 48 hours can induce differentiation to macrophages. Cells were seeded at a density of 1.5 × 105 cells per well on collagen coated 12-well plates and maintained at 37°C in a 5% CO2 atmosphere. Lipopolysaccharide (LPS; 100 ng/ml) were used to induce foam cell formation, inflammatory substances and cell apoptosis. Cell viability was determined by Trypan Blue. Murine macrophages were differentiated from bone marrow progenitors from the femurs of gene- knockout mice by M-CSF conditioned DMEM/10% FBS for 7 days [28 (link)].
4
Differentiation of THP-1 Monocytes into Macrophages
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THP-1 human monocytic leukemia cells (American Type Culture Collection, Manassas, VA, USA) were cultured in RPMI 1640 (Gibco-BRL, Grand Island, NY, USA), supplemented with 10% heat-inactivated fetal bovine serum (Gibco), 100 μg/mL penicillin, 100 μg/mL streptomycin, 10 mM HEPES, 1×MEM vitamin, and 0.5 µM 2-mercaptoethanol (Gibco). Cells were incubated at 37 °C in humidified air with 5% CO2. THP-1 cells at a density of 1 × 106/mL were differentiated into macrophages using 50 ng/mL phorbol 12-myristate 13-acetate (Sigma-Aldrich, St. Louis, MO, USA) for 72 h (THP-1 macrophages). Macrophage differentiation from monocytes was detected by their adherence to the culture plate.
5
Investigating Hcy-Induced Foam Cell Formation
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THP-1 human monocytic leukemia cells were obtained from the American Type Culture Collection (Rockville, MD, USA) and were maintained in RPMI 1640 supplemented with 10% FBS at 37°C in a humidified atmosphere of 5% CO2. Differentiation of THP-1 cells into macrophages was induced by using 100 nmol/L PMA for 24 h until adherent to the cell culture plate. To mimic the foam cell formation, we replaced the medium with 50 μg/mL ox-LDL in a serum-free medium for 24 h to fully differentiate THP-1 macrophages to foam cells. As described previously (10 (link)), we treated THP-1 macrophages with various concentration of Hcy (50, 100, and 200 μmol/L) or vehicle (equivalent volume of PBS) for 24 or incubated with 100 μmol/L Hcy under different exposure times (0, 6, 12, 24, and 48h). The concentration of Hcy was based on the previous studies (19 (link)–21 (link)). To explore the potential role of PCSK9, we first incubated THP-1 cells with different concentrations of SBC-115076 (5, 10, and 20 μmol/L) for 24 h to inhibit PCSK9 expression. Then we treated THP-1 cells together with Hcy (100 μmol/L) and SBC-115076 (20 μmol/L) for 24 h. We also incubated THP-1 macrophages with Hcy (100 μmol/L) and T0901317 (5 μg/mL) for 24 h. All cell experiments were based on a cell viability of 90% or higher. Each independent experiment was performed at least three times.
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