characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-06-18T17:02:48Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-08-21T18:21:43Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104221600
characteristics.characteristic: External Id
characteristics.text: UOM
characteristics.characteristic: INSDC center name
characteristics.text: 2018-06-18T17:02:48Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-08-21T18:21:43Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1880618
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5998:J_S10
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: mesothelial cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000077
characteristics.characteristic: cell type
characteristics.text: Norway rat
characteristics.characteristic: common name
characteristics.text: Protocols: Omental cells, from female Wistar rats weighing 220-250g (Charles River, Cambridgeshire, UK) were liberated by trypsin digestion (0.25% Trypsin-EDTA; Sigma-Aldrich, Dorset, UK) for 20min at 37°C. All subsequent procedures were carried out on ice. Cells were incubated with anti-mesothelioma (HBME-1) primary IgM antibody (Dako, Cambridge, UK. 1:50 for 30min) in PBS, 3% BSA, similar to previously described [PubMed: 27403660]. Cells were washed with wash buffer (PBS, 10% foetal calf serum, FCS) and incubated with biotinylated secondary antibody (Vector Laboratories, Peterborough, UK. 1:100 for 30min) before further wash steps. Cells at a concentration of 1.5x106/80μl were incubated with Streptavidin-coated nanobeads (Biolegend, London, UK, 1:5) 15min. The cell suspension was then made up to a volume of 3ml with wash buffer and placed in the Mojo magnet (Biolegend) for 5min. Unconjugated cells were poured off and the sorted population re-suspended in cell culture media consisting of DMEM high glucose supplemented with 15% FCS, 4mM L-glutamine (Sigma-Aldrich), 1% v/v penicillin/streptomycin and 0.4μg/ml hydrocortisone (Sigma-Aldrich). Female Wistar rats weighing 220-250g (Charles River, Cambridgeshire, UK) were housed with free access to food and water under specific pathogen free conditions at the University of Manchester. Temperature 20-22 °C, Humidity 40-60%, Specified rodent pathogen free conditions, 12 hours light/dark cycle. Nutrients Rat and Mouse Standard Diet no.1 expanded (Ban Kingman). Sorted mesothelial cells were seeded at 5x104/cm2 in multi-well plates. Adherent cells were apparent 48-72h post culture, at which point cells were washed with HBSS. To induce MMT, cells were placed in 5% serum containing media for 24h before exposure to 1ng/ml TGFβ1 (R&D systems, Abington, UK). Cells were collected in RNA protect (Thermo Fisher Scientific, Runcorn, UK) and RNA extracted using the RNeasy Plus Mini Kit (Qiagen). Truseq stranded mRNA Assay
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: IJ
characteristics.characteristic: individual
characteristics.text: Rattus norvegicus
characteristics.characteristic: organism
characteristics.text: Rattus norvegicus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10116
characteristics.characteristic: organism
characteristics.text: omentum
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0003688
characteristics.characteristic: organism part
characteristics.text: Rattus norvegicus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000383
characteristics.characteristic: sex
characteristics.text: Wistar
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001342
characteristics.characteristic: strain
characteristics.text: J_S10
characteristics.characteristic: title