|
| Status |
Public on Nov 05, 2023 |
| Title |
MDMpSTR_3 |
| Sample type |
RNA |
| |
|
| Source name |
MDM from donor 3 co-cultured in the presence of MM-BMSC
|
| Organism |
Homo sapiens |
| Characteristics |
individual: donor 3
disease state: healthy
cell type: MDM from healthy donor
protocol: co-cultured in the presence of MM-BMSC
|
| Treatment protocol |
After 24 hours cells were harvested by pipetting and stained with the 7-color mAb combination - HLADR-PacB, CD45-OC515, CD36-FITC, SLAN-PE, CD16-PECy7, CD300e-APC, CD14-APCH7 for FACSorting.
|
| Growth protocol |
MACS sorted CD14+ monocytes were differentiated into M2-like macrophages in RPMI supplemented with 10% FBS and M-CSF (10 ng/ml) for 7 days at 37ºC 5% CO2. Differentiated MDM were co-cultured for 24 hours in the presence of MM cells (1:5 ratio), in the presnece of confluet monolayer of MM-BMSC or both.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The cells were harvested with TRIzol Reagent (Invitrogen) and the RNA was extracted according to the manufacturer's instructions. As a last step of the extraction procedure, the RNA was purified with the RNeasy Mini-kit (Qiagen, Hilden, Germany).
|
| Label |
Biotin
|
| Label protocol |
The sense cDNA was prepared from 300 ng of total RNA using the Ambion® WT Expression Kit. The sense strand cDNA was then fragmented and biotinylated with the Affymetrix GeneChip® WT Terminal Labeling Kit (PN 900671).
|
| |
|
| Hybridization protocol |
Labeled sense cDNA was hybridized to the Affymetrix HuGene-2_0-st chip according to the the manufacturer protocols and using GeneChip® Hybridization, Wash and Stain Kit.
|
| Scan protocol |
Genechips were scanned with the Affymetrix GeneChip® Scanner 3000.
|
| Description |
Gene expression data from MDM cells from donor 3 co-cultured in the presence of MM-BMSC
|
| Data processing |
Data was normalized using Bioconductor software and RMA algorithm.
|
| |
|
| Submission date |
Nov 05, 2018 |
| Last update date |
Nov 05, 2023 |
| Contact name |
Aintzane Zabaleta |
| E-mail(s) |
azabaletaa@unav.es
|
| Organization name |
FOUNDATION FOR APPLIED MEDICAL RESEARCH
|
| Street address |
Avda. Pío XII 55
|
| City |
Pamplona |
| State/province |
Navarra |
| ZIP/Postal code |
31008 |
| Country |
Spain |
| |
|
| Platform ID |
GPL16686 |
| Series (1) |
| GSE122160 |
Expression data from monocyte-derived macrophages co-cultured in the presence of multiple myeloma (MM) cell line and/or MM-bone marrow stromal cell (BMSC) |
|
