|
| Status |
Public on Jan 28, 2013 |
| Title |
precursor-0day-rep1, miRNA expression |
| Sample type |
RNA |
| |
|
| Source name |
monocytes, rep1
|
| Organism |
Homo sapiens |
| Characteristics |
developmental stage: monocytes from PBMC
|
| Treatment protocol |
To generate immature DC (iDCs), monocytes were cultured for 1-6 days in the presence of 1000U/mL GM-CSF and 500 U/mL rhIL-4, followed by additional 1-2 days stimulation with 10ng/mL TNF-α, 10ng/mL IL-1β, 10ng/mL IL-6 and 10μg/mL PGE2 to get fully differentiated and matured DCs (mDCs). Medium exchange was performed every 2 days with fresh cytokine cocktails.
|
| Growth protocol |
Peripheral blood mononuclear cells (PBMC) were isolated from buffy coat by centrifugation at 2200rpm for 20min on lymphocyte separation medium. The phase containing white cells was removed and washed in RPMI-1640.CD14+ monocytes were separated with CD14+ microbeads and a magnetic separation column over a magnetic field (Miltenyi Biotec) according to the manufacture’s protocol. Eluted cells were washed thoroughly to remove the nonspecific binding to the beads. Purified monocytes were used as dendritic cell (DCs) precursors and were cultured in six-well plate in RPMI-1640 medium supplemented weih 10% heat inactivated fetal bovine serum (FBS), 2mM L-glutamine, 1Mm sodium pyruvate, 100U/mL penicillin and 100μg/mL streptomycin at 1 × 106 cells/mL at 37℃ in a 5% CO2 humidified incubator for different days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
microRNA samples were isolated from precursor, iDCs and mDCs using the mirVana RNA isolation kit according to the manufacturer’s protocol (Ambion). The microRNA quality, concentration, and integrity were analyzed using Agilent 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
According to the Agilent miRNA Microarray Protocol (Agilent Technologies).
|
| |
|
| Hybridization protocol |
According to the Agilent miRNA Microarray Protocol (Agilent Technologies).
|
| Scan protocol |
According to the Agilent miRNA Microarray Protocol (Agilent Technologies).
|
| Description |
miRNA expression of monocytes purified from PBMCs
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent Technologies). GeneView data were also analyzed by Feature Extraction Software.
|
| |
|
| Submission date |
Feb 27, 2009 |
| Last update date |
Jan 28, 2013 |
| Contact name |
Na Zhao |
| Organization name |
Kunming institute of zoology.CAS
|
| Street address |
Eastern Jiaochang road,32#
|
| City |
kunming |
| ZIP/Postal code |
650223 |
| Country |
China |
| |
|
| Platform ID |
GPL8227 |
| Series (2) |
| GSE15036 |
microRNA expression signature of monocyte-derived dendritic cells (DCs) from peripheral blood mononuclear cells (PBMCs) |
| GSE15076 |
Characterization of monocyte-derived dendritic cells (DCs) from peripheral blood mononuclear cells (PBMCs) |
|
