|
| Status |
Public on May 20, 2015 |
| Title |
Myostatin_24hr_300_3 |
| Sample type |
RNA |
| |
|
| Source name |
hSKMCs
|
| Organism |
Homo sapiens |
| Characteristics |
time: 24hr
dose: 300 ng/mL
treatment: Myostatin
|
| Treatment protocol |
human primary muscle cells 24 hours after stimulation with 10, 30 or 300 ng/ml of Myostatin, GDF11 and control (vehicle alone)
|
| Growth protocol |
Human skeletal muscle derived cells (hSkMDC; Cook Myosite) were cultured in growth medium consisting of MyoTonic™ basal medium (Cook Myosite) supplemented with 20% FBS (Hyclone) and MyoTonic™ serum free growth supplement (Cook Myosite). Differentiation was initiated 24 h after seeding (day 0) by changing to serum-free differentiation medium consisting of MyoTonic™ differentiation medium (Cook Myosite) containing 2% horse serum (Hyclone) and 1% FBS, and altering the atmospheric conditions from 5 to 7.5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the snap-fozen tibialis anterior muscle using an RNeasy Fibrous Tissue Mini Kit (Qiagen), and eluted in 40μl RNase free water. RNA concentration and quality was measured by a ND-1000 spectrophotometer (NanoDrop, Thermo Scientific). In case RNA quality was not acceptable (ratio 260/230<1.5), samples were cleaned using a RNeasy Plus Micro Kit (Qiagen), eluted in 15 μl and the new concentration and ratios measured again. RNA final quality was assessed on an Agilent 2100 Bioanalyzer using the Agilent RNA 6000 Nano kit, and processed for microarray when RIN>7
|
| Label |
biotin
|
| Label protocol |
Nugen Ovation RNA Amplification System+ WB + FL cDNA Biotin Module V2
|
| |
|
| Hybridization protocol |
Affymetrix Hybridization control kit (including B2)
|
| Scan protocol |
Affymetrix standard protocol
|
| Data processing |
RMA normalization followed by 2% 150 trimmed mean scaling. Probesets with scaled RMA values below 50 in all groups were filtered out
|
| |
|
| Submission date |
Mar 26, 2015 |
| Last update date |
May 20, 2015 |
| Contact name |
Sophie Brachat |
| E-mail(s) |
[email protected]
|
| Organization name |
Novartis Institutes For Biomedical Research
|
| Department |
Muskuloskeletal
|
| Street address |
Novartis Campus
|
| City |
Basel |
| ZIP/Postal code |
CH-4002 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE67326 |
GDF11 is a myokine that inhibits muscle differentiation and induces atrophy during regeneration |
|
