|
| Status |
Public on Jun 17, 2013 |
| Title |
Memory CD4 S + SN027 at 24h |
| Sample type |
RNA |
| |
|
| Source name |
purified CD45RA-CD4+ T cells from a healthy donor (+ 24h stimulation & treatment with SN from breast cancer patient 027)
|
| Organism |
Homo sapiens |
| Characteristics |
source tissue: peripheral blood
cell type: purified CD45RA-CD4+ T cells
disease state: healthy
treatment: + 24h stimulatin (anti-CD3 + anti-CD28) & treatment with SN from breast cancer patient 027
|
| Treatment protocol |
Cells (2x106 cells/ml) were incubated in X-VIVO 20 (Lonza) overnight following isolation. Cell viability was controlled the following day by flow cytometry (>95%) and then an equal volume of tumor SN (undiluted) or X-VIVO was added ± anti-CD3 (#317304, clone OKT3; Biolegend) and anti-CD28 (#555725, clone CD28.2; BD Pharmingen), each at 1µg/ml.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 100ng total RNA (3' IVT Express Kit Manual, 2009, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Human Genome U133 Plus 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner.
|
| Description |
Gene expression data from memory CD4+ T cells from healthy donor blood stimulated and treated with SN for 24h
Memory CD4+ T cells were negatively purified from a healthy donor (D6) blood using Memory CD4+ T Cell Isolation Kit (Myltenyi Biotec).
|
| Data processing |
The data (CEL files) were analyzed in R version 2.10.1 for RMA normalization.
|
| |
|
| Submission date |
Mar 23, 2012 |
| Last update date |
Sep 01, 2016 |
| Contact name |
Chunyan GU |
| E-mail(s) |
chunyan.gu@bordet.be
|
| Phone |
+32(0)25413711
|
| Organization name |
Free University of Brussels
|
| Department |
Medecine (Jules Bordet Institute)
|
| Lab |
Molecular Immunology
|
| Street address |
Bld de Waterloo 127
|
| City |
Brussels |
| ZIP/Postal code |
1000 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL570 |
| Series (2) |
| GSE36766 |
Effect of human breast tumor supernatant on normal CD4+ T cells (Confirmation Set SN) |
| GSE36769 |
CD4+ TIL in human breast cancer |
|
| Relations |
| Reanalyzed by |
GSE86362 |
