|
| Status |
Public on Mar 15, 2014 |
| Title |
Primary hepatocytes control |
| Sample type |
RNA |
| |
|
| Source name |
Primary hepatocytes
|
| Organism |
Homo sapiens |
| Characteristics |
cells: Primary hepatocytes
strain: PXB
treatment: none
|
| 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..
|
| |
|
| 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.
|
| 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.
|
| |
|
| 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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