|
| Status |
Public on Jan 29, 2009 |
| Title |
Dendritic cells after LPS treatment, time point 8 hours |
| Sample type |
RNA |
| |
|
| Source name |
monocyte-derived dendritic cells 8 hours after stimulation with LPS
|
| Organism |
Homo sapiens |
| Characteristics |
healthy donor
|
| Treatment protocol |
Day 5-immature DC were seeded on 24-well plates at 1×106 cells/ml with GM-CSF and IL-4. DC maturation was induced by addition of either a cocktail of pro-inflammatory cytokines and prostaglandins including TNF-alpha (10 ng/ml), IL-1-beta (10 ng/ml), IL-6 (1000 U/ml) and PGE2 (1 µg/ml), LPS (1 µg/ml), Poly(I:C) (50 µg/ml) or flagged CD40L (500 ng/ml) + anti-Flag mAb (1 µg/ml) for 4, 8 and 16 hours.
|
| Growth protocol |
Immature DC were generated from Buffy coats of healthy donors obtained from the German Red Cross after informed consent. Peripheral blood mononuclear cells (PBMC) were separated by Ficoll-Paque gradient centrifugation and DC were generated from purified CD14+ cells by using antibody-coated microbeads and magnetic separation . Selected CD14+ cells were cultured for 5 days at a concentration of 2×106/ml in the presence of GM-CSF (500 U/ml) and IL-4 (100 U/ml) (both purchased from Strahtmann Biotec, Hamburg, Germany).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from frozen cell pellets using the High Pure RNA Isolation kit according to the manufacturer’s (Roche) protocol.
|
| Label |
biotin
|
| Label protocol |
Total RNA was amplified and labelled using Message AmpII-Biotin enhanced Kit, Ambion
|
| |
|
| Hybridization protocol |
Following fragmentation, 12 ug of cRNA were hybridized for 16 hr at 45C on GeneChip U133 Plus 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450 using the EukGE-WS_v5_450 protocol
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000.
|
| Description |
Gene expression data from monocyte-derived dendritic cells 8 hours after stimulation with LPS
|
| Data processing |
The data were analyzed with GeneChip Operating Software version 1.4.0.036 (MAS 5.0) using Affymetrix default analysis settings. Normalization was performed by GCRMA.
|
| |
|
| Submission date |
Jan 29, 2008 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Elke Krueger |
| Organization name |
Charité Universitaetsmedizin Berlin
|
| Department |
Institute of Biochemistry
|
| Street address |
Monbijoustr. 2
|
| City |
Berlin |
| ZIP/Postal code |
10117 |
| Country |
Germany |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE10316 |
Monocyte-derived dendritic cells stimulated with LPS, Poly(I:C), CD40L or a combination of IL-1-b, IL-6, TNF-a and PGE2 |
|
| Relations |
| Reanalyzed by |
GSE49910 |
| Reanalyzed by |
GSE86362 |
| Reanalyzed by |
GSE119087 |
