Project description:Male and female muscle primary cultures were obtained from healthy young individuals. Cultures were treated with either estrogen, testesterone or DMSO. Gene expression was measured as response to the treatment
Project description:Dystrophin was knocked down in primary muscle cultures prepared from C57Bl/10 neonate mice using siRNA targeting dystrophin. Also in parallel, primary muscle cultures treated with siRNA targting luciferase were used as controls. The experiment was designed so that the same cell populations were used for both test and control conditions. This helped avoid the heterogeneity associated with the previous studies where test and control cell populations were non-identical. The experiment resulted in a clear transcriptome of dystrophin deficiency and demonstrated dystrophin as a major organizer of myogensis. Moreover, several interesting experimental targets were identified which can potentially open new lines of investigations.
Project description:Dystrophin was knocked down in primary muscle cultures prepared from C57Bl/10 neonate mice using siRNA targeting dystrophin. Also in parallel, primary muscle cultures treated with siRNA targting luciferase were used as controls. The experiment was designed so that the same cell populations were used for both test and control conditions. This helped avoid the heterogeneity associated with the previous studies where test and control cell populations were non-identical. The experiment resulted in a clear transcriptome of dystrophin deficiency and demonstrated dystrophin as a major organizer of myogensis. Moreover, several interesting experimental targets were identified which can potentially open new lines of investigations. 18 primary muscle cell cultures were prepared on 4 occasions. 7 cultures were treated with siRNAs targeting dystrophin, 7 received siRNA targeting firefly GL2 luciferase (treatment controls) and 4 remained untreated (untreated controls). RNAs were extracted 48 hours after differentiation induction for expression profiling analysis.
Project description:This experiment was aimed at understanding the transcriptomic response of primary muscle cell cultures to 24h rIL-1beta stimulation
Project description:In this experiment we have analyzed the effect of TGFbeta-1 incubation (3h of incubation time) on the gene expression profiles of human primary cultured skeletal muscle cells, both in growing myoblasts or in differentiated muscle fibres (after 10-days of differentiation), obtained from skeletal muscle biopsies from a 2-months old healthy donor and a 5-years old DMD (Duchenne Muscular Dystrophy) patient. All human skeletal muscle biopsies were obtained by surgery from calf muscles with informed consent and approval of the Human Use Committee of Vall d'Hebron University Hospital (Barcelona, Spain). Human myoblasts were cultured on 6 cm plates until reached 70-80% of confluence and then, were incubated for 3h in the presence or in absence of TGF-?1 (2.5 ng/ml). Skeletal muscle fibres were obtained by growing human myoblasts on 6 cm plates until reached 70-80% of confluence and then, cells were induced to differentiate by deprivation of growth factors in the culture medium. After 10 days of differentiation, skeletal muscle fibers were incubated for 3h in the presence or in absence of TGFbeta-1 (2.5 ng/ml). In all cases, immediately after TGFbeta-1 treatment the RNA was extracted and used for Affymetrix microarrays analysis using the U133 2.0 microarray genechips.
Project description:The goal was to examine and compare the transcriptomic profiles of primary muscle cell cultures (80% confluence) isolated from skeletal muscle (m. semitendinosus) derived from bulls belonging to different breeds and showing different levels of performance.
Project description:Vascular mineralization is a carefully orchestrated process, regulated by a number of promoters and inhibitors that function to ensure effective hydroxyapatite formation. Here we sought to identify new regulators of this process through a time series microarray analysis of mineralising primary vascular smooth muscle cell cultures over a 9 day culture period.