|
| Status |
Public on Feb 03, 2014 |
| Title |
Donor O, time 3.5 h |
| Sample type |
RNA |
| |
|
| Source name |
primary human monocytes stimulated with CCL2 for 2 h plus LPS for 1.5 h plus TNF-α for 0.5 h
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: early inflammatory monocytes
group: treated
|
| Treatment protocol |
The human primary blood monocytes were exposed in culture to sequential changes of microenvironmental conditions (chemokines and cytokines, temperature, bacterial-derived molecules, etc.) for 48 h.
|
| Growth protocol |
Human monocytes were obtained from buffy coats of healthy donors by isolating PBMC on Ficoll-Paque PLUS gradients and subsequent separation with Monocyte Isolation kit II. Monocytes were cultured in RPMI 1640+Glutamax-I Medium (GIBCO, Life Technologies) with 5% human AB serum, and gentamicine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA was performed using Qiagen miRNeasy kit (Qiagen) according to the manufacturer’s protocol
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared using GeneChip® 3’ IVT Express kit according to manufacturer's intructions
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Human Genome HG-U133 Plus 2.0 arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
Standard Affymetrix scanning procedures on a GCS3000 7G Affymetrix scanner operated by GCOS, GeneChip Operating Software
|
| Description |
Gene expression data from Donor O, time 3.5h
|
| Data processing |
RMA of .CEL files using affy Bioconductor library. In RMA, PM values have been background adjusted, normalized using quantile normalization, and expression measure calculated using median polish summarization
|
| |
|
| Submission date |
May 21, 2013 |
| Last update date |
Feb 03, 2014 |
| Contact name |
Silvio Bicciato |
| E-mail(s) |
silvio.bicciato@unipd.it
|
| Phone |
+39-049-827-6108
|
| Organization name |
University of Padova
|
| Department |
Molecular Medicine
|
| Street address |
via U. Bassi 59/b
|
| City |
Padova |
| ZIP/Postal code |
35131 |
| Country |
Italy |
| |
|
| Platform ID |
GPL17180 |
| Series (1) |
| GSE47122 |
Transcriptomic profiling of the development of the inflammatory response in human monocytes in vitro |
|
