|
| Status |
Public on Mar 01, 2020 |
| Title |
EVs no stimulation |
| Sample type |
RNA |
| |
|
| Source name |
Human immortalized cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: THP-1
treatment: EV
|
| Treatment protocol |
EVs were isolated with Total Exosome Isolation reagent (from serum)(Thermo Fisher Scientific), according to manufacturer´s intructions. To transfer EVs to cells, collected EVs from serum (100μL) were susupended with PBS (50μL) and incorporated to each well. As control PBS (50μL) was added to each well. Treated cells were cultured for 48 hours, stimulated with LPS (0.1mg/ml) and collected 2 hours later.
|
| Growth protocol |
THP-1 monocytes and THP-1 macrophages were cultured in RPMI-1640 with exosome free FCS media (5%). THP-1 monocytes were seeded onto 24-well plate and differentiated into macrophages with PMA (60ng/ml) for 16 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from cultured cells was extracted using TRIZOL (Invitrogen), according to the manufacturer´s instruction.
|
| Label |
Cy5
|
| Label protocol |
cDNA and Amino Allyl aRNA was synthesis by Amino Allyl MessageAmp II aRNA Amplification Kit (Ambion#1753). CyeDye Coupling and fragmentation were performed as the protocol supplied by TORAY Industries, Inc..
|
| |
|
| Hybridization protocol |
Hybridized for 16 h at 37 C with rotary shake (250 rpm). Hybridization buffer and washing protocol was followed by the protocol supplied by TORAY Industries, Inc..
|
| Scan protocol |
3D-Gene Scanner ((Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction(Toray Industries Inc., Tokyo, Japan).
|
| Description |
RNA was extracted from THP-1 cells cultured in exosome free medium supplemented with collected EVs, and was subjected to the microarray analysis.
|
| Data processing |
The raw data of each spot was normalized by substitution with a mean intensity of the background signal determined by all blank spots’ signal intensities of 95% confidence intervals.
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| |
|
| Submission date |
Mar 01, 2019 |
| Last update date |
Mar 02, 2020 |
| Contact name |
Satoshi Kondo |
| Organization name |
Toray Industries,Inc.
|
| Street address |
10-1 tebiro 6-chome
|
| City |
Kamakura |
| State/province |
Kanagawa |
| ZIP/Postal code |
248-8555 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13915 |
| Series (1) |
| GSE127714 |
mRNA expression profile in THP-1 macrophages treated with human serum extracellular vesicles |
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