|
| Status |
Public on Dec 18, 2007 |
| Title |
high DC strenght 2 |
| Sample type |
RNA |
| |
|
| Source name |
FACS-purified CFSE^lo CCR7+ IL-7R^ hi cells
|
| Organism |
Homo sapiens |
| Characteristics |
Total CD4+ T cells were isolated from the CD14- fraction by negative selection with a CD4+ T cell isolation kit on an Automacs (Miltenyi). CD4+ T cells were stained with anti-CCR7 mAb (R&D systems) followed by a biotin-labeled anti-g2a antibody and labeled streptavidin, FITC-labeled anti-CD45RA and PC5-labeled anti-CD8, anti-CD16 (Beckmann Coulter) and anti-CD19 (Becton Dickinson) and naïve CD4+ T cells (CD45RA+CCR7+) were purified by cell sorting to >99%. Then, T cells were DC stimulated, as desribed below
|
| Treatment protocol |
104 CFSE-labeled naïve T cells were co-cultured with TSST-pulsed 10^4 DCs
|
| Growth protocol |
co-cultured with TSST-pulsed DC in 200 microl /well complete medium in 96-round bottom wells. After 7 days CFSElo cells were purified on a FACSAria (Becton Dickinson) according to CCR7 and IL-7Ra expression (IL-7Ra mAbs, Beckmann coulter and R&D system).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
cells were lysed using Trizol (Invitrogen). An additional RNA clean-up step using the RNAeasy purification kit (Qiagen).
|
| Label |
biotin
|
| Label protocol |
Affymetrix GeneChip Expression 3’ Amplification One-Cycle Target Labeling kit from about 2ug of total RNA
|
| |
|
| Hybridization protocol |
standard Affymetrix procedures, washes performed using Fluidics Station 400
|
| Scan protocol |
standard Affymetrix procedures using a GeneChip Scanner 3000
|
| Description |
This sample is also known as IOSI06R-105
|
| Data processing |
Affymetrix GCOS 1.4
|
| |
|
| Submission date |
Dec 19, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Francesco Bertoni |
| E-mail(s) |
francesco.bertoni@ior.usi.ch
|
| Phone |
+41 91 820 0367
|
| Organization name |
IOR
|
| Department |
Lymphoma & Genomics Research Program
|
| Street address |
Via V. Vela 6
|
| City |
Bellinzona |
| State/province |
Ticino |
| ZIP/Postal code |
6500 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE6566 |
Strength of T cell stimulation |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE86362 |
| Reanalyzed by |
GSE119087 |
