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C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate

ABSTRACT: In urodele amphibians, limb regeneration involves the dedifferentiation of muscle myotubes into single cells that may acquire pluripotent potential. We have employed small molecules (myoseverin and BIO) to attempt to reproduce this behavior in mammalian muscle culture. C2C12 myotubes derived from the C2C12 myoblast cell line were induced to undergo cellularization by myoseverin treatment, which destabilizes tubulin filaments. The GSK-3 inhibitor, BIO, was then used to induce dedifferentiation. Induce neuron formation; the cells were incubated with 250 nM reversine for 48 h, and neural induction media (DMEM/F12 supplemented with N2 (Invitrogen)) and 1.5 uM all-trans retinoic acid for 7 days. C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate (derived by 20 M myoseverin treatment for 48 h) C2C12 myoblasts were differentiated into myotubes with 2%horse serum in DMEM for 8 days (from 2-4 d, 10 uM AraC treatment was also used to kill any remaining myoblasts). Next, myotubes were cellularized by 20 uM myoseverin treatment for 48 h. 24 h after myoseverin treatment, myotubes were treated with 10 uM BIO for 2d.

ORGANISM(S): Mus musculus

SUBMITTER: Darren Williams

PROVIDER: E-GEOD-34238 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate

Project description:In urodele amphibians, limb regeneration involves the dedifferentiation of muscle myotubes into single cells that may acquire pluripotent potential. We have employed small molecules (myoseverin and BIO) to attempt to reproduce this behavior in mammalian muscle culture. C2C12 myotubes derived from the C2C12 myoblast cell line were induced to undergo cellularization by myoseverin treatment, which destabilizes tubulin filaments. The GSK-3 inhibitor, BIO, was then used to induce dedifferentiation. Induce neuron formation; the cells were incubated with 250 nM reversine for 48 h, and neural induction media (DMEM/F12 supplemented with N2 (Invitrogen)) and 1.5 uM all-trans retinoic acid for 7 days. C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate (derived by 20 M myoseverin treatment for 48 h)

2011-12-08 | GSE34238 | GEO

Mus musculus

Project description:C2C12 murine myoblast cell line and 48 h 10 uM BIO treated C2C12 cellulate

| PRJNA149805 | ENA

Triggering acute RyR1 Ca2+ leak leads to improved mitochondrial remodelling and function

Project description:C2C12 myotubes at day 7 of differentiation were stimulated with a single session of SIT (6 x 30s with 4 min at rest) and were immediately after the stimulation treated or not with 10 uM S107 drug for 72h. The myotubes were then harvested at 72h post stimulation (for SIT condition) and 72h post stimulation and S107 treatment (for SIT S107 condition) for mass spectrometry analysis using the proteomics approach. N = 5 independent myotube wells per condition.

2021-10-20 | PXD018409 | Pride

Genome-wide analysis of gene expression during differentiation in C2C12 cells

Project description:Investigate the genome-wide gene expression profiles of 50% and 95% confluent C2C12 myoblasts and C2C12 myotubes differentiated for 24 and 48 hours.

2010-01-27 | GSE20059 | GEO

Gene expression profile of E1A infected C2C12 myotubes

Project description:Proliferating C2C12 myoblasts were induced to differentiate into myotubes and then infected with adenovirus expressing E1A (Ad-E1A), which induces cell cycle re-entry and dedifferentiation. We analyzed the transcriptional profile of E1A infected C2C12-myotubes through the Affymetrix Mouse Genome 430 2.0 Array, searching for genes that were significantly regulated between two independent biological replicates at two different time points (24h and 36h after infection with Ad-E1A). In addition, we took advantage of the E1A mutant known as YH47/dl928 (hereafter referred as YH47), which bears two mutations in the pocket-binding region of E1A (Y48H, C124G) able to disrupt the interaction with Rb and its cognate proteins and to impair cell-cycle re-entry phenotype. YH47 mutant was used to identify the Rb independent transcriptional reprogramming of C2C12. C2C12 cells were differentiated in vitro to myotubes as previously described. Myotubes were, then, infected with an adenovirus carrying the 12S form of E1A (dl520), with the YH47 E1A mutant (dl928) or with a control adenovirus (CTR) expressing a deletion of essentially the entire E1A gene (dl312). Two different time points after infection were considered (24 hours and 36 hours) to evaluate changes in C2C12 cells expression profile. Technical (A or B) and biological replicates (EXP1 or EXP2) were done for each condition.

2012-05-30 | E-GEOD-28457 | biostudies-arrayexpress

Gene expression profile of E1A infected C2C12 myotubes

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CoCl2 induced cytotoxicity is associated with dysregulated lncRNAs expression

Project description:To investigate differentially expressed lncRNAs in C2C12 myotubes with/without CoCl2 treatment, we used mouse lncRNA microarray to examine the expression of lncRNAs in C2C12 myotubes and C2C12 myotubes with CoCl2 treatment.

2021-05-28 | GSE115089 | GEO

CoCl2 induced cytotoxicity is associated with dysregulated circRNAs expression

Project description:To investigate differentially expressed circRNAs in C2C12 myotubes with/without CoCl2 treatment, we used mouse circRNA microarray to examine the expression of circRNAs in C2C12 myotubes and C2C12 myotubes with CoCl2 treatment.

2021-05-28 | GSE115088 | GEO

Transcription profiling of mouse C2C12 myotubes treated overnight with 500 uM palmitate or 1% bovine serum albumin control to identify mediators of obesity-linked reductions in PGC-1

Project description:To identify mediators of obesity-linked reductions in PGC-1, we tested the effects of cellular nutrients in C2C12 myotubes. While overnight exposure to high insulin, glucose, glucosamine, or amino acids had no effect, saturated fatty acids (FA) potently reduced PGC-1a and b mRNA expression. Experiment Overall Design: Cell culture - Mouse C2C12 myoblasts (ATCC, Manassas, VA) were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 20% fetal bovine serum (Invitrogen), at a confluency of 60-70%. To initiate differentiation, cells were allowed to reach 100% confluency and medium was changed to DMEM containing 2% horse serum (Invitrogen) and changed every 2 days. Full differentiation, with myotube fusion and spontaneous twitching, was observed at 5 days. Fatty acid stock preparation - Fatty acids were dissolved in 0.1 N sodium hydroxide (final concentration 100 mM) at 65 degrees C for 2 hours and then complexed with 10% fatty acid-free BSA, yielding a final stock of 5 mM. Three replicates for each fatty-acid. Microarray analysis - RNA was isolated as described from C2C12 myotubes treated overnight with 500 uM palmitate or 1% BSA, and cRNA was synthesized. 10 mg of cRNA were hybridized to Affymetrix mouse 430A 2.0 arrays. Intensity values were quantified using MAS 5.0 software. MAPPFinder (www.genmapp.org) was used to integrate expression data with known pathways. Transcription profiling of mouse C2C12 myotubes treated overnight with 500 uM palmitate or 1% BSA to identify mediators of obesity-linked reductions in PGC-1.

2007-11-15 | E-GEOD-6766 | biostudies-arrayexpress

Glucocorticoid receptor (GR) target genes in C2C12 myotubes

Project description:Analysis of C2C12 myotubes treated with dexamethasone (Dex) for 6 or 24 hours. Dex is a synthetic glucorticoid receptor agonist. Results provide insight to the effect of glucocorticoids on myotubes. C2C12 myotubes were cultured in DMEM supplemented with 2% horse serum, and treated with 1 u? Dex or an equal volume (0.01% v/v of media) of vehicle control ethanol for 6 or 24 hours. Total cellular RNA was isolated utilizing the NucleoSpin RNA II kit (Macherey-Nagel). RNA isolates were first quantified by standard spectrophotometry, and then qualitatively evaluated by capillary electrophoresis employing the Bio-Rad Experion system per manufacturer’s instruction. The final labeled cRNA samples were hybridized overnight to Illumina mouseWG-6 BeadChip arrays, which was performed at UCSF Genomic Core. All treatments were done in triplicates and the same batch of microarrays were used for all treatments. The Illumina expression arrays were pre-processed using lumi package. The differential expression analysis was performed using the Limma package.

2011-04-25 | E-GEOD-28840 | biostudies-arrayexpress

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