|
| Status |
Public on Jun 29, 2022 |
| Title |
FIS 343 3D exosomes-2R |
| Sample type |
RNA |
| |
|
| Source name |
Exosomes from FIS 343 primary culture grown in tumorspheres formation (3D) replicate 2
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Lung cancer cell line
cell type: FIS 343
histology: LUAD (lung adenocarcinoma)
|
| Growth protocol |
Primary culture cells were maintained in Dulbecco's Modified Eagle Medium/F12™ supplemented with 1% L-glutamine (Gibco-BRL Life Technologies), 10% fetal bovine serum (Gibco-BRL Life Technologies) and 1% penicillin/streptomycin (Gibco-BRL Life Technologies) at 37°C in a humidified atmosphere of 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cell lines were maintained in RPMI-1640 medium supplemented with MEM Non-Essential Amino Acids (0.001%) (Gibco-BRL Life Technologies), 10% fetal bovine serum (Gibco-BRL Life Technologies) and 1% penicillin/streptomycin (Gibco-BRL Life Technologies) at 37°C in a humidified atmosphere of 5% CO2.
Exosomes were isolated using ultracentrifugation from cell cultures supernatant and the samples were characterized in size, morphology and specific surface markers determination.
Total RNA was extracted from exosomes using Total RNA Purification Kit (Norgen Biotek, Canada) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
cRNA is converted to biotinylated single-stranded cDNA prepared according to the standard Affymetrix protocol from 13 ngs total RNA (GeneChip Whole Transcript WT Expression Arrays, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 5,5 ug of fragmented and labeled ss-cDNA were hybridized for 16 hr at 45ºC at 60 rpm on Hybridization Oven 640 de Affymetrix. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneArray Scanner 7G de Affymetrix
|
| Data processing |
Expression data were normalized, background and batch corrected using the Signal Space Transformation-Robust Multi-Chip Analysis (SST-RMA) implemented in the Transcriptome Analysis Console software version 4.0 (TAC 4.0).
|
| |
|
| Submission date |
Mar 09, 2022 |
| Last update date |
Jun 29, 2022 |
| Contact name |
Cristian Suarez Cabrera |
| E-mail(s) |
[email protected]
|
| Phone |
+34 699044222
|
| Organization name |
CIEMAT
|
| Lab |
Molecular Oncology Unit
|
| Street address |
Av. Complutense 40
|
| City |
Madrid |
| State/province |
Madrid |
| ZIP/Postal code |
28040 |
| Country |
Spain |
| |
|
| Platform ID |
GPL23126 |
| Series (1) |
| GSE198238 |
Analysis of exosomal cargo provides accurate clinical, histologic and mutational information in non-small cell lung cancer. |
|
