The human MUTZ-3 cell line was purchased from DSMZ (Germany). The cells were grown in α-MEM with 20% heat-inactivated FBS (PAA Laboratories-GE Healthcare Life Sciences), 1% Glutamax (Life Technologies) and 40 ng/ml GM-CSF (CellGro) as previously described (Nelissen et al., 2009 (link); Song et al., 2015 (link)). The MUTZ-3 cells were differentiated to immature DC with 62.5 ng/ml GM-CSF, 100 ng/ml IL-4 (CellGro) and 2.5 ng/ml TNF-α (CellGro) for 4 days (Zavašnik-Bergant and Bergant Marušič, 2016 (link)). The viability of the cells was evaluated using Trypan Blue (Sigma-Aldrich). The differentiated MUTZ-3 cells were matured with 20 ng/ml LPS (Sigma-Aldrich) for 24 h. Alternatively, the MUTZ-3 cells were stimulated with LPS in the presence of tick cystatin OmC2 (0.4 or 0.8 μM). Cystatin solution was filtered through a 0.22 μM Durapore PVDF membrane/Millex GV filter unit (Millipore). In control experiments, sterile PBS was added instead of cystatin OmC2, and the cells were cultured for 24 h. In addition, immature DC were matured with LPS in the presence of 0.4 or 0.8 μM human cystatin C (Zavašnik-Bergant et al., 2005 (link)).
Tnf α
Manufactured by Corning
Sourced in United States
TNF-α is a cytokine produced by various cells, including macrophages, lymphocytes, and endothelial cells. It plays a crucial role in the regulation of immune cells and is involved in the inflammatory response. This product is used for research purposes to study the function and activity of TNF-α in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Glutamax, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Trypan blue, by Merck Group (1 mentions)
Mutz 3 cell line, by Leibniz Institute DSMZ (1 mentions)
Gm csf, by Corning (1 mentions)
Interleukin 4 (il 4), by Corning (1 mentions)
Infliximab, by Johnson & Johnson (1 mentions)
Remicade, by Johnson & Johnson (1 mentions)
12 well plate, by Corning (1 mentions)
Heparin, by Leo pharma (1 mentions)
Tnf α, by Boehringer Ingelheim (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Corning (1 mentions)
24 well plate, by Corning (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Enbrel, by Pfizer (1 mentions)
5 protocols using tnf α
1
MUTZ-3 Dendritic Cell Maturation
2
Inflammatory Stimuli Effects on Keratinocytes
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Keratinocyte monolayer cultures and HEEs were stimulated with human IL‐1β (c: 100 ng/ml), TNFα (c: 10 ng/ml) ( both CellGro, Corning, NY) or TSLP (c: 10 ng/ml) (R&D systems, Minneapolis, MN) for 6 h or 24 h. Anakinra (Kineret, r‐metHuIL‐1ra, BoehringerIngelheim, Vienna, Austria) at a concentration of 50 µg/ml or Infliximab (Remicade, JanssenBiotech, Leiden, Netherlands) at a concentration of 100 µg/ml were added to the medium of HEEs 24 h and right before SDS application.
3
Endothelial Cell Response to TNF-α
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To determine the effect of TNF-α on the pro-inflammatory response of endothelial cells, HUVEC was cultured under static conditions in 12-well plates (Corning, Amsterdam, The Netherlands) and starved in fetal calf serum (FCS)-free EGM-2 BulletKit medium for 1 h. As control plasma, pooled plasma from five healthy humans was purchased from Innovative Research (IPLAK2E, Le-Perray-en-Yvelines, France). HUVEC was exposed to FCS-free medium containing 20% control plasma, 1 IU/ml heparin (Leo Pharma, Amsterdam, The Netherlands) and TNF-α (0.25–100 pg/ml, Boehringer Ingelheim, Ingelheim am Rhein, Germany) for 3 h. Thereafter, the medium was replaced by EGM-2 BulletKit medium for 1 h. The TNF-α concentrations used in vitro are comparable to concentration ranges reported for patient plasma during and after CABG surgery with CPB.[16 (link)]
4
Cytokine Exposure Effects on Apoptosis
5
Induction of IL-32 in Thyroid Cancer Cells
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To test whether induction of IL-32 mRNA expression in TPC-1 was caused by paracrine factors from the co-culture medium, fresh TPC-1 cells were stimulated with supernatants from TPC-1-monocyte co-culture experiments for 24 h. Since TNFα is an important inducer of IL-32 [1, (link)4] (link) and TNFα is abundantly present in the co-culture medium [21] (link), TPC-1 cells were also stimulated with TNFα (100 ng/ml R&D Systems, Minneapolis, MN, USA) for 24 h. Briefly, 500.000 cells/well were seeded in a 24-well plate (Corning, New York, USA) and left to adhere for 18 H. Medium was replaced with fresh medium mixed with supernatants from co-culture experiments in a 1:1 ratio or fresh medium with TNFα (100 ng/ml). To test whether IL-32 could also be induced in other TC cell lines, two additional cell lines were included, i.e., BC-PAP (papillary TC, BRAF V600E mutation) and FTC-133 (follicular TC, PTEN deficient) [22] (link). To confirm the role of TNFα, specific TNFα inhibitors, etanercept (decoy receptor, 10 μg/ml, Enbrel©, Pfizer, New York, USA) and adalimumab (monoclonal antibody, 10 μg/m, Humira©, Abbott GmbH & Co. Wiesbaden, Germany) were added to pre-selected experiments.
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