|
| Status |
Public on Nov 01, 2019 |
| Title |
M+IgG [Donor122_2] |
| Sample type |
RNA |
| |
|
| Source name |
monocyte derived dendritic cells
|
| Organism |
Homo sapiens |
| Characteristics |
condition: M+IgG
tissue: blood
time point: H4
|
| Treatment protocol |
Monocyte-derived DCs were pre- treated for one hour with mouse IgG1 (20 µg/mL, R&D Systems), mouse anti-IL10R blocking antibody (10 µg/mL, R&D Systems) or mouse anti-TNFα Receptors 1 and 2 (10 µg/mL, R&D Systems) and then cultured with medium or LPS (100 ng/mL, LPS-EB Ultrapure, activates TLR4 only, Invivogen) for additional 4 or 8 hours based on the time point
|
| Growth protocol |
Fresh blood samples collected from healthy donors were obtained from Hôpital Crozatier Établissement Français du Sang (EFS), Paris, France, in conformity with Institut Curie ethical guidelines. PBMCs were isolated by centrifugation on a Ficoll gradient (Ficoll-Paque PLUS, GE Healthcare Life Sciences). Monocytes were selected using antibody-coated magnetic beads and magnetic columns according to manufacturer’s instructions (CD14 MicroBeads, Miltenyi Biotec). To generate immature DCs, CD14+ cells were cultured for 5 days with IL-4 (50 ng/mL) and GM-CSF (10 ng/mL) in RPMI 1640 Medium, GlutaMAX (Life Technologies) with 10% FCS.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNeasy micro kit (Qiagen) in accordance with manufacturer's
|
| Label |
biotin
|
| Label protocol |
Samples were amplified and labelled according to the protocol recommended by Affymetrix for hybridization to Human Genome U133 Plus 2.0 arrays.
|
| |
|
| Hybridization protocol |
Samples were amplified and labelled according to the protocol recommended by Affymetrix for hybridization to Human Genome U133 Plus 2.0 arrays.
|
| Scan protocol |
Affymetrix GeneTitan device ( combined hybridization oven, fluidics processing, and imaging device )
|
| Description |
gene expression data for MoDC after 4 hours in medium + isotype ctrl
|
| Data processing |
Expression data were normalized with Plier. Transcriptomics analysis was performed in a Matlab environment. For independent filtering, we used the function geneverfilter, which calculates the variance of each probe across the samples and identifies the ones with low variance. Probes with variance less the 40th percentile were filtered out because poorly informative.
|
| |
|
| Submission date |
Oct 31, 2016 |
| Last update date |
Dec 17, 2020 |
| Contact name |
Vassili Soumelis |
| Organization name |
INSERM
|
| Department |
U976 - HIPI
|
| Lab |
Human systems immunology and inflammatory networks
|
| Street address |
1 avenue Claude Vellefaux
|
| City |
Paris |
| ZIP/Postal code |
75010 |
| Country |
France |
| |
|
| Platform ID |
GPL17692 |
| Series (1) |
| GSE89342 |
Dissection of intercellular communication using the transcriptome-based framework ICELLNET |
|
