|
| Status |
Public on Jun 30, 2012 |
| Title |
Placebo group, 12 months, patient number 34 |
| Sample type |
RNA |
| |
|
| Source name |
Isolated PBMCs from diabetic male patient_Placebo_12 months
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
dietary group: placebo (A)
time point: 12 months
cell type: Isolated peripheral blood mononuclear cells (PBMCs)
|
| Treatment protocol |
PBMCs were isolated at day 0, day 180 (6months) and day 365 (12 months) from the following groups:i) placebo (control), n=6 arrays (6 diabetic males); ii) grape extract, n=6 arrays (6 diabetic males); iii) resveratrol-enriched grape extract, n=6 arrays (6 diabetic males)
|
| Growth protocol |
Male patients' heparinised blood was processed within two hours after extraction and used to isolate PBMCs. Isolation was carried out under sterile conditions to avoid the activation of monocytes. Blood was diluted 1:1 with RPMI 1640 cell culture medium and processed by density gradient centrifugation with Histopaque-1077 (Sigma Aldrich) according to the manufacturer instructions. After washing the cells (x2) with phosphate buffer solution (PBS), the PBMCs were lysed using RLT Buffer (Qiagen) and stored at –80C for RNA extraction.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using AllPrep DNA/RNA/Protein Mini Kit (Qiagen) according to the manufacturer's instructions. RNA was eluted in RNAse free water, aliquoted and stored at –80C.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the One-Cycle target IVT labeling Affymetrix protocol from 4 μg total RNA (Expression Analysis Technical Manual, 2004, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 18 cRNA were hybridized for 16 hr at 45C on GeneChip Affymetrix Human Gene 1.0 ST. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000.
|
| Description |
A-S34_12months
|
| Data processing |
The data were analyzed with Robust Multichip Average (RMA), using CEL files obtained from Affymetrix Expression Console (version 1.1.2).
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| |
|
| Submission date |
Mar 29, 2012 |
| Last update date |
Jun 30, 2012 |
| Contact name |
María-Teresa García-Conesa |
| E-mail(s) |
mtconesa@cebas.csic.es
|
| Phone |
+ 34 968 396276
|
| Organization name |
CEBAS-CSIC
|
| Department |
Quality, Safety and Bioactivity of Plant Foods
|
| Street address |
Campus de Espinardo
|
| City |
Espinardo |
| State/province |
Murcia |
| ZIP/Postal code |
30100 |
| Country |
Spain |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE36930 |
Comparative transcriptomic profiling of peripheral blood mononuclear cells (PBMCs) isolated from patients in secondary prevention of CVD taking placebo, grape extract or resveratrol-enriched grape extract |
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