|
| Status |
Public on Aug 12, 2017 |
| Title |
Early differentiating myocytes from T2DM donor 4 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Early differentiating myocytes, T2DM donor
|
| Organism |
Homo sapiens |
| Characteristics |
disease state: T2DM
individual: T2 Diabetic subject 4
gender: male
age: 66
body composition: non-obese
tissue: vastus lateralis
cell type: Early differentiating myocytes
|
| Treatment protocol |
Cells were not treated
|
| Growth protocol |
Cells were grown in 20% FBS, 1% PS and 1% FZ in F10/HAM, in 1% matrigel coated culture dishes. At 100% confluent cells, cells were transferred to intermediate medium (DMEM containing 1 g/L glucose, 10% FBS and 1% PS). After 2 days, medium was changed to differentiation media (DMEM containing 4.5 g/L glucose, 2% horse serum and 1% PS) in order to induce differentiation into myotubes (myocytes)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Hy3
|
| Label protocol |
500 ng total RNA from sample and reference was labeled with Hy3™ and Hy5™ fluorescent label, respectively, using the miRCURY LNA™ microRNA Hi-Power Labeling Kit, Hy3™/Hy5™ (Exiqon, Denmark) following the procedure described by the manufacturer
|
| |
|
| Channel 2 |
| Source name |
Common reference pool (healthy subjects only)
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: isolated satellite cells
|
| Treatment protocol |
Cells were not treated
|
| Growth protocol |
Cells were grown in 20% FBS, 1% PS and 1% FZ in F10/HAM, in 1% matrigel coated culture dishes. At 100% confluent cells, cells were transferred to intermediate medium (DMEM containing 1 g/L glucose, 10% FBS and 1% PS). After 2 days, medium was changed to differentiation media (DMEM containing 4.5 g/L glucose, 2% horse serum and 1% PS) in order to induce differentiation into myotubes (myocytes)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Hy5
|
| Label protocol |
500 ng total RNA from sample and reference was labeled with Hy3™ and Hy5™ fluorescent label, respectively, using the miRCURY LNA™ microRNA Hi-Power Labeling Kit, Hy3™/Hy5™ (Exiqon, Denmark) following the procedure described by the manufacturer
|
| |
|
| |
| Hybridization protocol |
The hybridization was performed according to the miRCURY LNA™ microRNA Array instruction manual using a Tecan HS 4800™ hybridization station (Tecan, Austria)
|
| Scan protocol |
Slides were scanned using the Agilent G2565BA Microarray Scanner System (Agilent Technologies, Inc., USA) and the analyzed using the ImaGene 9.0 software (BioDiscovery, Inc., USA)
|
| Description |
T2 Diabetic subject 4 of 8
D-4-2
Raw files: 0_Exiqon_*.txt represents Hy3 signals, 1_Exiqon_*.txt represents Hy5 signals.
|
| Data processing |
LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Agilent software was used.
|
| |
|
| Submission date |
Aug 25, 2016 |
| Last update date |
Aug 12, 2017 |
| Contact name |
Søren Nielsen |
| E-mail(s) |
soren_nielsen@inflammation-metabolism.dk
|
| Organization name |
Rigshospitalet
|
| Department |
Section 7641
|
| Lab |
CFAS/CIM
|
| Street address |
Blegdamsvej 9
|
| City |
Copenhagen |
| State/province |
- |
| ZIP/Postal code |
2100 |
| Country |
Denmark |
| |
|
| Platform ID |
GPL22371 |
| Series (1) |
| GSE86069 |
Dysregulation of a miR-23b/27b-p53 axis impairs muscle differentiation in humans with type 2 diabetes |
|
