|
| Status |
Public on Dec 31, 2013 |
| Title |
CB70BS2 NT |
| Sample type |
RNA |
| |
|
| Source name |
Cord-blood derived monocytes
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Cord-blood derived monocytes
treatment: untreated
|
| Treatment protocol |
TEM were exposed for 2 hours to the treatments (100 ng/ml excepted TNF 20ng/ml), washed, kept for 30 h in culture and used for RNA extraction.
|
| Growth protocol |
TEM were differentiated in vitro from CD34+ cord-blood precursors using SCF (Stem Cell Factor), Interleukine-6, TPO (Thrombopoietin), Flt-3 ligand and MCF-7 conditioned medium as previously reported (Laurent et al., Cancer Res. 2011 Jun 1;71(11):3781-91)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs from 10 000 monocytes were isolated and purified with the Qiagen RNeasy micro plus extraction kit.
|
| Label |
biotin
|
| Label protocol |
For each sample, 100ng of total RNA were amplified using the WT sense strand Target Labelling kit (Affymetrix, Cat.no. 900223); 5.5 microg of the resulting sense cDNA was fragmented by UDG (uracil DNA glycosylase) and APE 1 (apurinic/apyrimidic endonuclease 1) and biotin-labelled with TdT (terminal deoxynucleotidyl transferase) using the GeneChip® WT Terminal labelling kit (Affymetrix Cat.no. 900671, Santa Clara, USA).
|
| |
|
| Hybridization protocol |
Affymetrix Human Gene 1.0 ST arrays (Affymetrix, Santa Clara , CA, USA) were hybridized with 2.7 microg of biotinylated target, at 45°C for 17 hours washed and stained according to the protocol described in Affymetrix GeneChip® Expression Analysis Manual (Fluidics protocol FS450_0007).
|
| Scan protocol |
The arrays were scanned using the GeneChip® Scanner 3000 7G (Affymetrix) and raw data was extracted from the scanned images and analyzed with the Affymetrix Power Tools software package (Affymetrix).
|
| Description |
Gene expression data from untreated human Cord-blood derived monocytes
|
| Data processing |
Background subtraction, RNA normalization and probeset summarization were performed using the Affymetrix Power Tools software package
|
| |
|
| Submission date |
Dec 19, 2011 |
| Last update date |
Dec 31, 2013 |
| Contact name |
Marie-Agnès Doucey |
| E-mail(s) |
Marie-Agnes.Doucey@unil.ch
|
| Phone |
+41216925960
|
| Organization name |
LICR@UNIL
|
| Street address |
Biopôle 3; Ch. des Boveresses 155
|
| City |
Epalinges |
| ZIP/Postal code |
1066 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL10739 |
| Series (1) |
| GSE34559 |
Tie-2 expressing monocytes (TEM) expression data |
|
