|
| Status |
Public on Dec 25, 2016 |
| Title |
mDC-BC2-TIV-09-6h-in vitro |
| Sample type |
RNA |
| |
|
| Source name |
mDC-TIV-09-6h-in vitro
|
| Organism |
Homo sapiens |
| Characteristics |
^sample: mDC_BC2_TIV-09_6h
cell population: mDC
donor: BC2
stimulus: TIV-09
timepoint: 6h
experiment: mDC vs TIV-09/MIV-09
sample type: in vitro
|
| Treatment protocol |
Whole blood samples were drawn by finger prick. DC samples were prepared as follows: Buffy coats obtained from healthy donors (Carters Blood Care, Dallas, TX) were fractionated on a Ficoll gradient. Peripheral blood mononuclear cells (PBMCs) were further enriched for DCs through negative selection using the Pan DC kit (Stemcell Technologies). The enriched cells were sorted on a FACS Aria (BD Biosciences) for LIN-/ HLA-DR+/ CD123+ and CD11c- pDCs and LIN- / HLA DR+ / CD123- and CD11c+ mDCs. Purity was routinely >98%. DCs were stimulated at a density of 5x104 cells per well with vaccines (6 µL/mL) in 200 µL complete medium in 96-well plates for 0, 6, 16, 24 and 48h. For microarray analysis, 0.5x106 cells were cultured with (6 µL/mL) vaccine in 0.5 mL complete medium for 6h
|
| Extracted molecule |
total RNA |
| Extraction protocol |
After vaccine stimulation, cells were lysed in RLT buffer (RNeasy Plus Mini Kit, Qiagen, Hercules, CA), 1% β-mercaptoethanol and stored at -80oC. Total RNA was isolated from cell lysates according to manufacturer’s instructions.
|
| Label |
biotin
|
| Label protocol |
RNA from samples passing quality control was amplified and labeled with Illumina TotalPrep RNA amplification kit (Illumina, San Diego, CA). RNA integrity was assessed using an Agilent 2100 Analyzer (Agilent Technologies, Palo Alto, CA).
|
| |
|
| Hybridization protocol |
Biotinylated complementary RNA (cRNA) was hybridized to Illumina Human-6 Beadchip Array version 2 using Illumina standard protocol
|
| Scan protocol |
Hybridized chips were scanned in Illumina Beadstation 500.
|
| Description |
5147861026_E
|
| Data processing |
Fluorescent hybridization signals were assessed with Beadstudio software (Illumina). Statistical analysis and hierarchical clustering were performed using GeneSpring 7.3.1 software (Agilent Technologies)
|
| |
|
| Submission date |
May 20, 2016 |
| Last update date |
Mar 16, 2023 |
| Contact name |
Nicole Baldwin |
| E-mail(s) |
Nicole.Baldwin@BSWHealth.org
|
| Organization name |
Baylor Research Institute
|
| Street address |
3434 Live Oak St
|
| City |
Dallas |
| ZIP/Postal code |
75204 |
| Country |
USA |
| |
|
| Platform ID |
GPL6884 |
| Series (2) |
| GSE81690 |
Influenza Vaccines Differentially Regulate the Interferon Response in Human Dendritic Cell Subsets [HumanWG-6 v3.0 Beadchip Experiments] |
| GSE81692 |
Influenza Vaccines Differentially Regulate the Interferon Response in Human Dendritic Cell Subsets |
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