|
| Status |
Public on May 21, 2016 |
| Title |
Co-culture between tumor cells and mMDSC (Lung 4T1) |
| Sample type |
RNA |
| |
|
| Source name |
co-culture
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Tumor cell+mMDSC (Lung 4T1)
|
| Treatment protocol |
Single cell suspensions were prepared from bone marrows, lungs and tumor tissues. Bone marrow cells were obtained by flushing bones with PBS using a 28G 1/2 syringe. Tumor and lung tissues were dissociated and digested with collagenase for 1hr at 37°C. Red blood cells were lysed by ACK lysis buffer. These cells were labeled with fluorescence-conjugated Ly6C, Ly6G and CD11b antibodies andsorted with a FACS Aria cell sorter. Sorted cells were lysed using RLT lysis buffer from Qiagen RNeasy Mini Kit.
|
| Growth protocol |
Murine breast cancer cells expressing the luciferase gene were implanted into the fat pads of 5-week –old BALB/c or C57BL/J6 mice.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Extraction of total RNA was performed with Qiagen RNeasy Mini Kit according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the standard Affymetrix protocol from 2 ng total RNA (GeneChip WT Pico Reagent kit, Affymetrix).
|
| |
|
| Hybridization protocol |
Following labeling, 5.5 ug of cDNA were hybridized for 16 hr at 45C on Mouse transcriptome 1.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G.
|
| Description |
Gene expression data from co-cultured tumor cells
|
| Data processing |
CEL files were processed with Gene Level-SST-RMA algorithm of the Expression Console 1.4. Differential expressions were calculated on Transcriptome Analysis Console, TAC (Affymetrix) using CHP files and CEL files.
|
| |
|
| Submission date |
May 20, 2016 |
| Last update date |
May 21, 2016 |
| Contact name |
Anne-Pierre Morel |
| Organization name |
Cancer Research Center of Lyon
|
| Street address |
28, rue Laennec, Cheney D 2nd floor
|
| City |
Lyon |
| State/province |
Rhone-Alpes |
| ZIP/Postal code |
69008 |
| Country |
France |
| |
|
| Platform ID |
GPL20775 |
| Series (1) |
| GSE81701 |
Gene expression data from MDSC subsets, cell lines and tumor cells |
|
