organism: Homo sapiens
description: Protocols: Samples were collected using Qiazol. Human tissue-derived myoblasts - Primary myoblasts were maintained in a myoblast medium: DMEM/F-12, HEPES (31330-038, Thermo Fisher Scientific) supplemented with 10 % fetal bovine serum (FBS, Hyclone, Logan, UT), 10 ng/mL fibroblast growth factor 2 (FGF2, 100-18B, Peprotech), and 50 nM Dexamethasone (D4902, Sigma-Aldrich) on 0.1 % gelatin (G1393, Sigma-Aldrich) coated culture ware. Human tissue-derived myotubes - Primary myoblasts were differentiated into myotubes. Cells were seeded at 600 cells/cm2 on 0.1 % gelatin coated cultureware in myoblast medium containing 1 mM Acid ascorbic 2P (A8960, Sigma-Aldrich). Human induced pluripotent stem cells - Primary myoblasts were reprogrammed into hiPSCs following the protocol described in Massourides et al., using the Yamanaka's factors POU5F1, SOX2 and KLF4 transduction by ecotropic or amphotropic vectors (Table S3). HiPSCs were adapted and maintained with mTeSR™1 culture medium (05850, Stemcell Technologies) on Corning® Matrigel® Basement Membrane Matrix, lactose dehydrogenase elevating virus (LDEV)-Free-coated cultureware (354234, Corning Incorporated). Cells were then seeded at 20,000 cells/cm2, passaged and thawed each time with 10 μM StemMACS™ Y27632. RNA extraction was done using the miRNeasy Mini kit (217004, QIAgen) on the QIAcube instrument. RNAs coming from the part B of the extraction protocol was used for miRseq. PartB RNA was quantified and purity/quality was assessed on the 2100 Agilent BioAnlayzer using the Agilent small RNA kit (5067-1548, Agilent). 10 ng of miRNA was reverse transcribed using the Ion Total RNA-seq kit v2 (4475936, Thermofisher Scientific) following the protocol of the manufacturer for small RNA libraries. The cDNA libraries were amplified and barcoded using Ion Total RNA-seq kit v2 and Ion Xpress RNA-seq Barcode Adapters 1-16 Kit (Thermofisher Scientific). The amplicons were quantified using Agilent High Sensitivity DNA kit before the samples were pooled in sets of fifteen.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-03-01T04:03:08Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-09-02T10:07:41Z
attributes.tag: External Id
attributes.value: SAMEA5928495
attributes.tag: INSDC center name
attributes.value: INSERM UEVE UMR861, I-STEM, AFM
attributes.tag: INSDC first public
attributes.value: 2020-03-01T04:03:08Z
attributes.tag: INSDC last update
attributes.value: 2019-09-02T10:07:41Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8293:D3_DMD Cell 03-1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: M418
attributes.tag: cell type
attributes.value: mesoderm
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: Duchenne muscular dystrophy
attributes.tag: genotype
attributes.value: Stop exon 7 c
attributes.tag: progenitor cell type
attributes.value: induced pluripotent stem cell
attributes.tag: sample name
attributes.value: E-MTAB-8293:D3_DMD Cell 03-1
attributes.tag: scientific_name
attributes.value: Homo sapiens