Project description:Peripheral blood was acquired from the Pécs Regional Blood Transfusion Center (Pécs, Hungary) and processed using Lymphocyte Separation Medium 1077 (Promocell, Heidelberg, Germany) in accordance with the manufacturer's instructions. Subsequently, the viable mononuclear cells were isolated, counted, and seeded into 6-well plates at a density of 1,000,000 cells per well. The culture medium was specifically formulated using RPMI-1640 medium (Biowest, Nuaillé, France), enriched with 1% L-glutamine (Lonza, Basel, Switzerland), 2% penicillin/streptomycin (Lonza, Basel, Switzerland), and 10% exosome-depleted FBS (Thermo Fisher Scientific, Waltham, USA) to minimize EV interference from bovine serum, further supplemented with 100 ng/mL Peprotech M-CSF 300-25 (Gibco, Waltham, USA). The cells were cultured for 5 days in a 5% CO2 incubator at 37 °C, resulting in the differentiation of all monocytes into macrophages (M0) (Cao et al., 2019).
2025-01-17 | GSE286937 | GEO
Project description:NDM-Producing Enterobacterales in Switzerland from 2019 to 2020
Project description:Peripheral blood was acquired from the Pécs Regional Blood Transfusion Center (Pécs, Hungary) and processed using Lymphocyte Separation Medium 1077 (Promocell, Heidelberg, Germany) in accordance with the manufacturer's instructions. Subsequently, the viable mononuclear cells were isolated, counted, and seeded into 6-well plates at a density of 1,000,000 cells per well. The culture medium was specifically formulated using RPMI-1640 medium (Biowest, Nuaillé, France), enriched with 1% L-glutamine (Lonza, Basel, Switzerland), 2% penicillin/streptomycin (Lonza, Basel, Switzerland), and 10% exosome-depleted FBS (Thermo Fisher Scientific, Waltham, USA) to minimize EV interference from bovine serum, further supplemented with 100 ng/mL Peprotech M-CSF 300-25 (Gibco, Waltham, USA). The cells were cultured for 5 days in a 5% CO2 incubator at 37 °C, resulting in the differentiation of all monocytes into macrophages (M0) (Cao et al., 2019). Differentiated macrophages were treated with MEV (Milk-derived extracellular vesicles) from healthy and mastitis groups separately to achieve the final concentration of 2×10^11 particles/mL. Additionally, untreated cell cultures with the same volume of PBS were maintained as negative controls. After 48 hours, macrophage supernatants were collected and stored at -80 °C (for a maximum of 3 months) for future experiments.
Project description:Peripheral blood was acquired from the Pécs Regional Blood Transfusion Center (Pécs, Hungary) and processed using Lymphocyte Separation Medium 1077 (Promocell, Heidelberg, Germany) in accordance with the manufacturer's instructions. Subsequently, the viable mononuclear cells were isolated, counted, and seeded into 6-well plates at a density of 1,000,000 cells per well. The culture medium was specifically formulated using RPMI-1640 medium (Biowest, Nuaillé, France), enriched with 1% L-glutamine (Lonza, Basel, Switzerland), 2% penicillin/streptomycin (Lonza, Basel, Switzerland), and 10% exosome-depleted FBS (Thermo Fisher Scientific, Waltham, USA) to minimize EV interference from bovine serum, further supplemented with 100 ng/mL Peprotech M-CSF 300-25 (Gibco, Waltham, USA). The cells were cultured for 5 days in a 5% CO2 incubator at 37 °C, resulting in the differentiation of all monocytes into macrophages (M0) (Cao et al., 2019). Differentiated macrophages were treated with MEV (Milk-derived extracellular vesicles) from healthy and mastitis groups separately to achieve the final concentration of 2×10^11 particles/mL. Additionally, untreated cell cultures with the same volume of PBS were maintained as negative controls. After 48 hours, macrophage supernatants were collected and stored at -80 °C (for a maximum of 3 months) for future experiments.
Project description:Using RNA sequencing to map differentially expressed genes in human brain microvascular endothelial cells challenged with DIII domain of protein E of WNV and DIII domain of protein E of TBEV.
Project description:Using RNA sequencing to map differentially expressed genes in the 3D model of the blood-brain barrier ( composed of human brain endothelial cells, human astrocytes, and human pericytes) challenged with TBEV.
Project description:Flavivirus non-structural protein 1 (NS1) is secreted from cells in infected individuals and in cell culture and levels correlate with disease severity. Treatment of murine bone marrow–derived dendritic cells (BMDCs) with recombinantly produced solube NS1 (sNS1) of tick-borne encephalitis virus (TBEV) or West Nile virus (WNV) prior to poly(I:C) stimulation revealed two gene clusters that were downregulated by TBEV or WNV sNS1 and that were associated with innate and adaptive immune responses. Especially co-stimulatory molecules and pro-inflammatory cytokines as well as chemokines were found to be downregulated in sNS1 pre-treated BMDCs prior to poly(I:C) stimulation.