characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2017-08-07T15:33:53Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-04-11T12:49:05Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA103973335
characteristics.characteristic: External Id
characteristics.text: University of Cambridge
characteristics.characteristic: INSDC center name
characteristics.text: 2017-08-07T15:33:53Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-04-11T12:49:05Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1662503
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5674:cR_FiPS_R1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: fibroblast derived cell line
characteristics.characteristic: cell line
characteristics.text: pluripotent stem cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0002248
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: Conventional hPSC culture: Human pluripotent stem cells, iPS cells and re-primed human naive stem cells were routinely maintained in KSR/FGF medium: DMEM/F-12 (Sigma-Aldrich, D6421) supplemented with 10 ng/ml FGF2 (prepared in-house), 20% KnockOut Serum Replacement (KSR) (Thermo Fisher Scientific), 100 mM 2-mercaptoethanol (2ME) (Sigma-Aldrich, M7522), 1X MEM nonessential amino acids (Thermo Fisher Scientific, 11140050) and 2 mM L-glutamine (Thermo Fisher Scientific, 25030024) on irradiated mouse embryonic fibroblasts (MEFs). Cells were passaged as clusters by detachment with dispase (Sigma-Aldrich, 11097113001). To establish PB-Oct4EOS stable transfectants, 1 ug/ml of puromycin was applied for two passages (10 days) to transfected cultures on Matrigel (Roche). Naïve stem cell culture: Chemically reset and embryo-derived (HNES) human naive stem cells were propagated in N2B27 supplemented with t2iLGo (1μM CHIR99021, 1μM PDO325901, 10ng/ml human LIF and 2uM Go6983) with or without Rho-associated protein kinase (ROCK) inhibitor (Y-27632), and plated either on MEFs or tissue laminin (Sigma). Where indicated as tt2iLGo, CHIR99021 was used at 0.3 uM. For feeder-free culture, Geltrex (growth factor-reduced) or laminin were added to the medium at time of plating. Cells were cultured in 5% O2, 7% CO2 in a humidified incubator at 37C and passaged by dissociation with Accutase (Thermo Fisher Scientific, A1110501) or TrypLE (Thermo Fisher Scientific, 12605028) every 3-5 days. Cells were cryopreserved in CryoStem medium (Biological Industries, K1-0640).
characteristics.characteristic: description
characteristics.text: tt2iLGo
characteristics.characteristic: growth condition
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: cR_FiPS_R1
characteristics.characteristic: title