Project description:Gene expression analysis in human skeletal myoblasts (undifferentiated mononucleated cells, cultured in growth medium - 15% FBS) and human skeletal myotubes (pre-formed myotubes, cultured in differentiation medium – 2% horse serum) exposed to the HDACi TSA (50nM) for 24 hours TSA treated undifferentiated human primary myoblasts and terminally differentiated human myotubes
Project description:Gene expression analysis in human skeletal myoblasts (undifferentiated mononucleated cells, cultured in growth medium - 15% FBS) and human skeletal myotubes (pre-formed myotubes, cultured in differentiation medium – 2% horse serum) exposed to the HDACi TSA (50nM) for 24 hours
Project description:Genome-wide gene expression analysis of MyoD-infected DMD-specific iPSCs (GM05112-M5.1) on days 0 (untreated), day 3 and day 8 post Dox treatment, human primary myoblasts (undifferentiated and as differentiated myotubes), and undifferentiated iPSCs from healthy donors (iPSCs-1 and iPSCs-2).
Project description:Genome-wide gene expression analysis of MyoD-infected DMD-specific iPSCs (GM05112-M5.1) on days 0 (untreated), day 3 and day 8 post Dox treatment, human primary myoblasts (undifferentiated and as differentiated myotubes), and undifferentiated iPSCs from healthy donors (iPSCs-1 and iPSCs-2). DMD-specific iPSCs were infected with lentivirus expressing MyoD under the control of Tet-inducible promoter and another lentivirus expressing the transactivator. To initiate myogenic differentiation, iPSCs were treated with 1µg/ml Dox. RNA was isolated 0, 3 and 8 days later and gene expression analysis was performed.
Project description:Maps of genomic regions in proximity to the nuclear lamina were determined in undifferentiated C2C12 myoblasts (MBs) and 6 day differentiated C2C12 myotubes (MTs) using DamID with a Dam-Lamin B1-encoding lentivirus.
Project description:Muscle cells are potential targets of many arboviruses, such as Ross River, Dengue, Sindbis, and Chikungunya viruses, that may be involved in the physiopathological course of the infection. During the recent outbreak of Zika virus (ZIKV), myalgia was one of the most frequently reported symptoms. We investigated the susceptibility of human muscle cells to ZIKV infection. Using an in vitro model of human primary myoblasts that can be differentiated into myotubes, we found that myoblasts can be productively infected by ZIKV. In contrast, myotubes were shown to be resistant to ZIKV infection, suggesting a differentiation-dependent susceptibility. Infection was accompanied by a caspase-independent cytopathic effect, associated with paraptosis-like cytoplasmic vacuolization. Proteomic profiling was performed 24h and 48h post-infection in cells infected with two different isolates. Proteome changes indicate that ZIKV infection induces an upregulation of proteins involved in the activation of the Interferon type I pathway, and a downregulation of protein synthesis. This work constitutes the first observation of primary human muscle cells susceptibility to ZIKV infection, and differentiation-dependent restriction of infection from myoblasts to myotubes. Since myoblasts constitute the reservoir of stem cells involved in reparation/regeneration in muscle tissue, the infection of muscle cells and the viral-induced alterations observed here could have consequences in ZIKV infection pathogenesis.
Project description:undifferentiated mouse skeletal muscle cells (myoblasts) and differentiated mouse skeletal muscle cells (myotubes) were subjected to treatment with siRNAs targeting SETD2 or a non-targeting siRNA (si-Ctrl). Differential gene expression analysis and splicing analysis was performed on the resulting data.
Project description:Human primary myoblasts were differentiated to myotubes and treated with 0, 10, 100, or 500ng/ml intelectin for 6 hours. Total RNA was extracted and profiled on Affymtrix Clariom D GeneChips reannotatwed to ENSG identifiers using the Brainarray V25 cdf.
Project description:hTERT/cdk4 immortalized myogenic human cell lines represent an important tool for skeletal muscle research, being used as therapeutically-pertinent models of various neuromuscular disorders and in numerous fundamental studies of muscle cell function. However, the cell cycle is linked to other cellular processes such as integrin regulation, the PI3K/Akt pathway, and microtubule stability, raising the question as to whether transgenic modification of the cell cycle results in secondary effects that could undermine the validity of these cell models. Here we subjected healthy and disease lines to intensive transcriptomic analysis, comparing immortalized lines with their parent primary populations in both differentiated and undifferentiated states, and testing their myogenic character by comparison with non-myogenic (CD56-negative) cells. We found that immortalization has no measurable effect on the myogenic cascade or on any other cellular processes, and that it was protective against the systems level effects of senescence that are observed at higher division counts of primary cells. This dataset includes gene expression profiles for 94 samples comprising primary myoblasts and their corresponding immortalized clones in both differentiated and undifferentiated states (average of 4 cell culture replicates each) from 5 human subjects (2 healthy and 3 Duchenne muscular dystropy - DMD), together with primary populations of non-myogenic (CD56-ve) cells from the muscles of 8 other human subjects. Total RNA was extracted from, myoblasts, myotubes (after 9 days of differentiation), or CD56-ve cells by dissolving cell pellets in TRIzol then using PureLink RNA Mini Kit.
