characteristics.text: SAMN31853179
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: 2023-02-02T04:38:24.020Z
characteristics.characteristic: INSDC last update
characteristics.text: SRS15853656
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: SRS15853656
characteristics.characteristic: SRA accession
characteristics.text: throphoblast stem cells derived from naive induced pluripotent 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 H9 naive-iPSCs
characteristics.characteristic: source_name
characteristics.text: H9-TSC_Rep_2
characteristics.characteristic: title
characteristics.text: Naive hPSCs did a pre-treatment on MEF with TS medium for 24h, whereupon naive hPSCs were single-cell dissociated by TrypLE Express, and 0.25-0.5 × 106 cells were seeded on a 12-well plate pre-coated with 5 μg/mL Collagen IV or on mitotically inactivated mouse embryonic fibroblasts and cultured in 0.8 mL TS medium. Cells were cultured in 5% CO2 and 5% O2, media was changed every 2 days, and passaged upon 80-100% confluency at a ratio of 1:2 to 1:4. During derivation from naive hPSCs, the cells grew slowly during the initial few passages. Between passage 5 and 10, highly proliferative hTSCs emerged.
characteristics.characteristic: treatment protocol