characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-04-01T00:17:00Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-04-01T00:17:00Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA12131272
characteristics.characteristic: External Id
characteristics.text: EMBL
characteristics.characteristic: INSDC center name
characteristics.text: 2022-04-01T00:17:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-04-01T00:17:00Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS9777883
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11334:Somitoids_S01
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Human iPS cells (201B7 with HES7 promoter-luciferase reporter and the fluorescent nuclear reporter)
characteristics.characteristic: cell line
characteristics.text: induced pluripotent stem cell
characteristics.characteristic: cell type
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: Generation of human somitoids with 10% Matrigel. Once human iPSCs reached 50-70% confluence, the cells were washed with PBS(-) twice and incubated with 2 ml 0.5 mM EDTA in PBS at 37ºC for 6-7 min. The cells were mechanically dissociated by pipetting and transferred into 8 ml of the N2B27 medium described below. The cell suspension was centrifuged at 152 xg for 3 min and resuspended into 10 ml of N2B27 medium. The washing step was repeated once, and the supernatant was completely removed. The cell pellet was resuspended into 150-500 μl of the somitoid induction medium described below with 10 µM Y27632. The cell concentration was calculated and adjusted to 7 cells/ml with an adequate amount of pre-warmed induction medium with Y27632. Then, 50 µl of the cell suspension (= 350 cells) was aliquoted into each well of a U-Shaped-Bottom, 96-well-plate (Nunclon Sphera 96U-well plate, Thermo Scientific, 174925) by using a multichannel pipette. To settle down the cells on the bottom of the plate, the 96-well-plate was centrifuged at 152 xg for 2 min before incubation. One day after aggregate formation, 150 µl of fresh somitoid induction medium without Y27632 was added to each well. On days 2 and 3, 150 µl of the medium was carefully removed and replaced with 150 µl of fresh N2B27 medium without disturbing the aggregate. On day 4, 150 µl of the medium was removed from the wells and replaced with 150 µl of fresh N2B27 medium containing 10% of growth factor reduced Matrigel (Corning, 356231). The medium was not changed after the Matrigel addition. On day 7, somitoids were collected. RNA extraction included in the single-cell nucleic acid library construction protocol cDNA libraries were constructed with Chromium Single Cell 3' GEM, Library & Gel Bead Kit v3 (10x Genomics) according to manufacturer's protocol.
characteristics.characteristic: description
characteristics.text: normal
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000461
characteristics.characteristic: disease
characteristics.text: With-Matrigel; +CHIR99021_8-10
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: Somitoids_S01
characteristics.characteristic: title