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 A of the extraction protocol was used for mRNA-seq. PartA RNA was quantified on Nanodrop spectrophotometer (ND-1000, Thermo Fisher Scientific) and purity/quality (RIN ≥ 7) was assessed on the 2200 TapeStation using the Agilent RNA ScreenTape (5067-5576 / 5067-5577 / 5067-5578, Agilent). Libraries are prepared with TruSeq Stranded mRNA kit protocol according supplier recommendations. Briefly, the key stages of this protocol are successively, the purification of PolyA containing mRNA molecules using poly-T oligo attached magnetic beads from 1µg total RNA, a fragmentation using divalent cations under elevated temperature to obtain approximately 300bp pieces, double strand cDNA synthesis and finally Illumina adapter ligation and cDNA library amplification by PCR for sequencing.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-03-01T04:03:08Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2019-09-23T13:21:44Z
attributes.tag: External Id
attributes.value: SAMEA5790605
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: 2019-09-23T13:21:44Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8321:D25_Healthy Cell 03-1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: M398
attributes.tag: cell type
attributes.value: induced pluripotent stem-derived cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: Embryonic/fetal skeletal muscle
attributes.tag: disease
attributes.value: normal
attributes.tag: replicate
attributes.value: 1
attributes.tag: sample name
attributes.value: E-MTAB-8321:D25_Healthy Cell 03-1
attributes.tag: scientific_name
attributes.value: Homo sapiens