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Status |
Public on May 02, 2014 |
Title |
human_dendritic cells_7day in DC culture |
Sample type |
RNA |
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Source name |
human monocyte derived cells
|
Organism |
Homo sapiens |
Characteristics |
in vitro culture: 7day in GMCSF/IL4 DC culture
|
Treatment protocol |
DCs were matured by 300ng/ml LPS (0111:B4, Sigma) on day 5 and harvested on day 7 for mature DCs.
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Growth protocol |
Monocytes were purified from PBMCs using anti-CD14 microbeads (Miltenyi Biotech) and then cultured at 37°C in 24-well plates (5x10^5 cells per well) in 1 mL of RPMI-1640 Medium (PAA) with 10% (vol/vol) FCS (PAA) containing 100 ng/mL human GM-CSF and 20 ng/mL human IL-4 (R&D Systems,Minneapolis, MN). Half medium was replaced by fresh medium with GM-CSF and IL-4 at day 3 and day 5.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated from cells at indicated time point with Trizol.
|
Label |
biotin
|
Label protocol |
Biotinylated cDNA were prepared according to the standard Affymetrix protocol from 250 ng total RNA by using Ambion® WT Expression Kit. WT Expression Kit
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|
|
Hybridization protocol |
Following labeling, 5.5 ug of cDNA were hybridized for 16 hr at 45℃ on GeneChip Human Transcriptome Array 2.0 in Hybridization Oven 645. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
Scan protocol |
GeneChips were scaned by using Affymetrix® GeneChip Command Console (AGCC) which installed in GeneChip® Scanner 3000 7G.
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Data processing |
The data were analyzed with Robust Multichip Analysis (RMA) algorithm using Affymetrix default analysis settings and global scaling as normalization method.
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Submission date |
Jan 16, 2014 |
Last update date |
May 02, 2014 |
Contact name |
PIN WANG |
E-mail(s) |
pinqi21sh@163.com
|
Organization name |
SMMU
|
Street address |
Xiangyin Road 800
|
City |
Shanghai |
ZIP/Postal code |
200433 |
Country |
China |
|
|
Platform ID |
GPL17586 |
Series (1) |
GSE54143 |
DC-specific lincRNA controls dendritic cell development and function through interacting with STAT3 |
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