|
| Status |
Public on Feb 19, 2018 |
| Title |
non-treated primary HBEC-derived EVs |
| Sample type |
RNA |
| |
|
| Source name |
Exosome
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HBEC
treatment: CSE (-)
isolated by ultracentrifugation: extracellular vesicles (EVs)
|
| Treatment protocol |
Cigarette smoke extract (CSE) was prepared for smoking exposure model in vitro. Forty milliliters of cigarette smoke was drawn into the syringe and slowly bubbled into sterile serum-free cell culture media in a 15-ml BD falcon tube. One cigarette was used for the preparation of 10 mL of solution. CSE solution was filtered (0.22 μm) to remove insoluble particles and was designated as a 100% CSE solution. We analyzed the role of CSE induced HBEC-derived EVs in the airway microenvironment. After HBECs were incubated in the non-FBS containing medium with or without a low-concentration CSE (1.0%) for 2 days, we collected conditioned medium (on day 4, 6) and isolated EVs by ultracentrifugation.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from EVs using QIAzol and the miRNeasy Mini Kit (Qiagen) according to the manufacturer’s protocol.
|
| Label |
Cy3
|
| Label protocol |
RNA sample was labeled with the miRNA Complete Labeling Reagent and Hybridization kit (Agilent Technologies) according to the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Labeled RNA was hybridized to the Human miRNA Microarray version 2 8x15K (G4470B Agilent Technologies) with the miRNA Complete Labeling Reagent and Hybridization kit (Agilent Technologies) according to the manufacturer’s instructions.
|
| Scan protocol |
Microarray slides were scanned in an Agilent Technologies G2505C Microarray Scanner at 5 micron resolution.
|
| Description |
Sample name: CSE (-)
Human bronchial epithelial cell
|
| Data processing |
Images were quantified using Agilent Feature Extraction software version 10.7.3.1 (Agilent) using default parameters (Grid: 046064_D_F_20121223) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Feb 19, 2015 |
| Last update date |
Feb 19, 2018 |
| Contact name |
Yu Fujita |
| Organization name |
National Cancer Center Research Institute
|
| Department |
Research of Molecular Functions and Targets
|
| Lab |
Division of Molecular and Cellular Medicine
|
| Street address |
5-1-1 Tsukiji
|
| City |
chuo-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
104-0045 |
| Country |
Japan |
| |
|
| Platform ID |
GPL18044 |
| Series (1) |
| GSE66119 |
MicroRNA microarray between smoking-induced human bronchial epithelial cell (HBEC)-derived extracellular vesicles (EVs) and non-treated HBEC-derived Evs |
|
