characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-10-01T16:26:41Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-10-01T16:26:41Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA111379498
characteristics: External Id
characteristics: Department of Dermatology, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland
characteristics: INSDC center name
characteristics: 2022-10-01T16:26:41Z
characteristics: INSDC first public
characteristics: 2022-10-01T16:26:41Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS13476481
characteristics: SRA accession
characteristics: E-MTAB-12199:B7_3
characteristics: Submitter Id
characteristics: 30
characteristics: year
characteristics: age
characteristics: ArrayExpress
characteristics: broker name
characteristics: T helper 9 cell
characteristics: cell_type
characteristics: human
characteristics: common name
characteristics: Protocols: Peripheral Blood Mononuclear Cells (PBMC) were isolated by Ficoll-Plaque Plus (GE Healthcare, UK) density gradient centrifugation. Human naïve CD4+ T Cell Isolation Kit Human (Stemcell Technologies) as per the manufacturer’s instructions. Naïve T cells were stimulated with anti-CD3/CD2/CD28 beads (T cell/bead = 2:1, Miltenyi) and primed into effector CD4+ T cell subsets with IL-12 (5 ng/ml) (BioLegend) for TH1 cells, IL-4 (50 ng/ml) (BioLegend) for TH2 cells, IL-4 (50 ng/ml) and TGF-b (5 ng/ml) (R&D Systems) for TH9 cells, and TGF-b (5 ng/ml) for iTREG. From cell culture initiation to analysis, the culture medium was supplemented with the indicated cytokines every other day. Culture medium consisted of PRMI-1640 with Hepes (GIBCO) supplemented with 5% of heat-inactivated human serum (Swiss Red Cross, Basel, Switzerland), 2mM L-Glutamine (Biochrom), 50U/ml penicillin and 50μg/ml streptomycin (Bioconcept) and 50 IU/ml IL-2 (Roche). T cells were cultured at a density of 0.25-1x105cells/96 well plate in a total volume of 200μL cell culture medium Naïve T cells were stimulated with anti-CD3/CD2/CD28 beads (T cell/bead = 2:1, Miltenyi) and primed into effector CD4+ T cell subsets with IL-12 (5 ng/ml) (BioLegend) for TH1 cells, IL-4 (50 ng/ml) (BioLegend) for TH2 cells, IL-4 (50 ng/ml) and TGF-b (5 ng/ml) (R&D Systems) for TH9 cells, and TGF-b (5 ng/ml) for iTREG. Total RNA was isolated with RNAeasy kit (Qiagen) Illumina TruSeq Stranded mRNA protocol (Directional, first strand)
characteristics: description
characteristics: adult
characteristics: developmental stage
characteristics: normal
characteristics: disease
characteristics: IL-4 and TGF-b
characteristics: growth condition
characteristics: B
characteristics: individual
characteristics: Homo sapiens
characteristics: organism
characteristics: Homo sapiens
characteristics: organism
characteristics: blood
characteristics: organism part
characteristics: Homo sapiens
characteristics: scientific_name
characteristics: male
characteristics: sex
characteristics: antibodies against CD3, CD2, and CD28 (7 days)
characteristics: stimulus
characteristics: B7_3
characteristics: title