|
| Status |
Public on Mar 15, 2014 |
| Title |
Primary hepatocytes HBV infected with miR93 expression |
| Sample type |
RNA |
| |
|
| Source name |
Primary hepatocytes
|
| Organism |
Homo sapiens |
| Characteristics |
cells: Primary hepatocytes
strain: PXB
treatment: HBV infected with miR93
|
| Treatment protocol |
When infection, 3 ml of the serum containing 9.0 log IU/ml of HBV genotype C, which is approximately 1.5 × 107 copies of HBV, was added for 7 days. Bionanocapsules containing miR93 was added at final concentration of 50 nM of the miRNA.
|
| Growth protocol |
Cells were cultured in a high density in DMEM containing 10% FBS, 5 ng/ml EGF, 0.25 µg/ml Insulin, 0.1 mM Ascorbic acid, and 2% DMSO in a 24-well plate.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Standard RNA extraction method using Trizol Reagent was used.
|
| 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..
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| |
|
| 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 |
Human primary hepatocytes were isolated from chimeric mice (PXB mice) and cultured in vitro. HBV from patinet's serum was infected and its replication was confirmed. At day 6, cells were treated with bionanocapsules containing miR93 for24h. Day7 after infection, cells were harvested and analyzed.
|
| 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. Measurements of both duplicate spots with the signal intensities greater than 2 standard deviations (SD) of the background signal intensity were considered to be valid. A relative expression level of a given mRNA was calculated by comparing the signal intensities of the averaged valid spots with their mean value throughout the microarray experiments.
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| |
|
| Submission date |
Mar 14, 2014 |
| Last update date |
Mar 17, 2014 |
| Contact name |
Satoshi Kondo |
| Organization name |
Toray Industries,Inc.
|
| Department |
New Projects Development Division
|
| Street address |
Tebiro 6-10-1
|
| City |
Kamakura |
| State/province |
Kanagawa |
| ZIP/Postal code |
248-8555 |
| Country |
Japan |
| |
|
| Platform ID |
GPL13915 |
| Series (1) |
| GSE55928 |
Changes of mRNA expression levels in HBV-infected human primary hepatocytes and those cells treated with bionanocapsules containing microRNA-93. |
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