|
Status |
Public on Aug 12, 2017 |
Title |
Late differentiating myocytes from T2DM donor 8 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
Late differentiating myocytes, T2DM donor
|
Organism |
Homo sapiens |
Characteristics |
disease state: T2DM individual: T2 Diabetic subject 8 gender: male age: 52 body composition: non-obese tissue: vastus lateralis cell type: Late 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
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|
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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 8 of 8 D-8-3 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.
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|
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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 |
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