|
| Status |
Public on Aug 09, 2010 |
| Title |
Control group, Time 0 (basal), male pig number 15 |
| Sample type |
RNA |
| |
|
| Source name |
PBMNCs, male pig, control group fed a normal chow, time 0 (basal expression levels)
|
| Organism |
Sus scrofa |
| Characteristics |
breed: Mini Pig
gender: male
tissue: blood
cell type: peripheral blood mononuclear cells (PBMNCs)
diet: normal
time: 0
pig: 15
|
| Treatment protocol |
PBMNCs were isolated from the following groups: i) CT (control group), time 0, n=10 arrays (5 females, 5 males; basal expression levels); ii) CT group, time 9 months, n=4 arrays (2 females, 2 males); iii) High-fat fed animals (H-F group), time 9 months, n=4 arrays (2 females, 2 males); iv) High-fat fed animals supplemented with resveratrol (H-F+RES group), time 9 months, n=4 arrays (2 females, 2 males).
CT diet: 5% beef tallow; H-F diet: 20% beef tallow; H-F+RES diet: 20% beef tallow plus 8 mg/Kg body weight per day resveratrol.
|
| Growth protocol |
Pigs' heparinised blood was processed within two hours after extraction and used to isolate PBMNCs. 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 PBMNCs were lysed using RLT Buffer (Qiagen) and stored at –80 ºC for RNA extraction.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using an RNeasy ® Plus Mini Kit (Qiagen) according to the manufacturer's instructions. RNA was eluted in RNAse free water, aliquoted and stored at –80ºC.
|
| 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).
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| |
|
| Hybridization protocol |
Following fragmentation, 15 cRNA were hybridized for 16 hr at 45C on GeneChip Affymetrix Porcine Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000.
|
| Description |
CT0.15.M
Gene expression data from PBMNCs isolated from male pig; control animal, time 0 (basal levels of expression).
|
| Data processing |
Using CEL files obtained from GCOS software (Affymetrix), the data were analyzed with Robust Multichip Average (RMA) using Affymetrix Expression Console (v.1.1) software.
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| |
|
| Submission date |
Aug 08, 2010 |
| Last update date |
Sep 17, 2010 |
| 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 |
GPL3533 |
| Series (1) |
| GSE23503 |
Comparative transcriptomic profiling of peripheral blood mononuclear cells (PBMNCs) isolated from pigs exposed to a high-fat diet (H-F) or to a high-fat diet plus resveratrol (H-F+RES) |
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