characteristics.text: SAMN31853174
characteristics.characteristic: External Id
characteristics.text: Department of Molecular Medicine, University of Padua
characteristics.characteristic: INSDC center name
characteristics.text: 2023-02-01T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-02-02T04:38:30.120Z
characteristics.characteristic: INSDC last update
characteristics.text: SRS15853650
characteristics.characteristic: INSDC secondary accession
characteristics.text: live
characteristics.characteristic: INSDC status
characteristics.text: Generic
characteristics.characteristic: NCBI submission model
characteristics.text: Generic.1.0
characteristics.characteristic: NCBI submission package
characteristics.text: SRS15853650
characteristics.characteristic: SRA accession
characteristics.text: human embryonic stem cells
characteristics.characteristic: cell type
characteristics.text: hTSCs were cultured in 12-well plate coated with 5 μg/mL Collagen IV (Corning, 354233) at 37°C overnight or on mitotically inactivated mouse embryonic fibroblasts. Cells were cultured in 0.8 mL TS medium [DMEM/F12 supplemented with 0.1 mM 2-mercaptoethanol, 0.2% FBS, 0.5% Penicillin-Streptomycin, 0.3%, BSA (Gibco 15260-037), 1% ITS-X (Gibco, 51500), 1.5 μg/ml L-ascorbic acid (Sigma A4544), 50 ng/ml EGF (peprotech AF-100-15), 2 μM CHIR99021 (Axon Medchem cat. nos 1386 and 1408), 0.5 μM A83-01 (Axon Medchem 1421), 1 μM SB431542 (Axon Medchem 1661), 0.8 mM VPA (HDACi, Sigma, P4543), and 5 μM Y-27632] and in 5% CO2 and 5% O2. Media were changed every 2 days, and cells were passed using TrypLE Express every 3-4 days at a ratio of 1:8.
characteristics.characteristic: growth protocol
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: TSC derived from KiPS primed-iPSCs
characteristics.characteristic: source_name
characteristics.text: ccTSC.01_p10_Rep_3
characteristics.characteristic: title
characteristics.text: Chemical resetting from primed to naive PSC. Cells were seeded at 10000 cells/cm2 on mitotically inactivated MEFs in E8 medium with 10µM ROCKi (added only for 24 hours). Two days after plating (day 0), medium was changed to PDL/HDACi (N2B27 with 1µM PD0325901, 10ng/ml human LIF and 1mM VPA (HDACi)). Following 3 days in PDL/HDACi, medium was changed to PXGL for a further 10-11 days before passing in PXGL medium on MEFs (see Fig. 1a). For the chemical resetting from primed to ccTSCs, KiPS were seeded at 10000 cells/cm2 on mitotically inactivated MEFs in E8 medium. Two days later (day 0) medium was changed to PDL/HDACi. Following 3 days in PDL/HDACi, medium was changed to PXGL.
characteristics.characteristic: treatment protocol