|
| Status |
Public on Oct 31, 2011 |
| Title |
GB-associated stromal cells 3 |
| Sample type |
RNA |
| |
|
| Source name |
brain
|
| Organism |
Homo sapiens |
| Characteristics |
disease state: Glioblastoma (GB)
cell type: stromal cell
|
| Growth protocol |
Before cell isolation, we removed meninges and visible blood vessels from the biopsy specimens, which were then minced and mechanically dissociated in Dulbecco’s modified Eagles’ medium (DMEM) (Lonza, Verviers, Belgium). Aliquots of the resulting suspension corresponding to samples before culture were stored at -80°C for further analysis by real-time quantitative PCR (Q-PCR) and flow cytometry analysis of DNA. The remainder of the suspension was used to seed T80 flasks (Nunc, Dominique Dutsher, Brumath, France) containing DMEM supplemented with 10% human AB serum (HABS) (EFS, Lyon, France) and 1% antibiotics (Sigma-Aldrich, Saint Quentin Fallavier, France). Cells were grown at 37°C in a humidified incubator, under an atmosphere containing 5% CO2, and the medium was changed twice weekly. Primary cultures were used under passage 10.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using NucleoSpin RNAII Kit (Macherey-Nagel)
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 350 ng RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
Hybridizations were performed following the manufacturer's recommendations (protocol version 5.7)
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B) using one color scan setting for 4x44k array slides
|
| Description |
GASCs 3
Agilent Feature Extraction file:
251485048430_SLOT01_S01_GE1_105_Dec08_1_3.txt
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters
|
| |
|
| Submission date |
Sep 13, 2010 |
| Last update date |
Oct 31, 2011 |
| Contact name |
Amandine Etcheverry |
| Organization name |
CNRS UMR 6290 - IGDR
|
| Street address |
2 rue Henri Le Guilloux
|
| City |
Rennes |
| ZIP/Postal code |
35033 |
| Country |
France |
| |
|
| Platform ID |
GPL4133 |
| Series (1) |
| GSE24100 |
Isolation and characterization of cancer-associated fibroblast-like cells in the glioblastoma microenvironment |
|
