|
| Status |
Public on Aug 11, 2017 |
| Title |
Hepatogenic differentiation, Hepatocyte |
| Sample type |
protein |
| |
|
| Source name |
Hepatocytes
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Hepatocytes
strain: Balb/C
Sex: male
|
| Treatment protocol |
See growth protocol
|
| Growth protocol |
Mouse MSCs were cultured in an mMSC-maintenance medium, low glucose DMEM supplemented with 10% FBS; mouse primary hepatocytes were cultured in Hep-maintenance medium, and DMEM/F12 supplemented with 10% FBS; HD 28 cells are differentiated from mMSCs by following methods. MSCs were treated with a Step-1 differentiation medium, consisting of IMDM supplemented with 20 ng/ml HGF and 10 ng/ml bFGF (R&D Systems), and nicotinamide 0.61 g/L (Sigma-Aldrich), for 7 days, then treated with a Step-2 differentiation medium, consisting of IMDM supplemented with 20 ng/ml oncostatin M (Invitrogen), 1 μmol/L dexamethasone (Sigma-Aldrich), and 50 mg/ml ITS+ premix (BD Biosciences). The medium was changed once per week to generate HD 28 cells. dHD 28 cells are derived from HD 28 cells by following methods. Hepatogenic de-differentiation was induced in HD 28 cells derived from MSCs by replacing the Step-2 medium with MSC-maintenance medium.
|
| Extracted molecule |
protein |
| Extraction protocol |
Cell lysate were lysed in lysis buffer containing protease and phosphatase inhibitors (Sigma-Aldrich) according to the manufacturer’s directions.
|
| Label |
Streptavidin-conjugated fluorescent dye (Cy3 equivalent)
|
| Label protocol |
Protocol is following the manufacturer’s directions (Ray biotech Mouse L308 Array, Glass Slide Cat# AAM-BLG-1-4). See http://www.raybiotech.com/files/manual/Antibody-Array/AAM-BLG-1.pdf
|
| |
|
| Hybridization protocol |
Protocol is following the manufacturer’s directions (Ray biotech Mouse L308 Array, Glass Slide Cat# AAM-BLG-1-4). See http://www.raybiotech.com/files/manual/Antibody-Array/AAM-BLG-1.pdf
|
| Scan protocol |
Protocol is following the manufacturer’s directions (Ray biotech Mouse L308 Array, Glass Slide Cat# AAM-BLG-1-4). See http://www.raybiotech.com/files/manual/Antibody-Array/AAM-BLG-1.pdf
|
| Description |
raw file: Ray biotech Mouse L308 Array(block1=Heps block2=dHD28).xlsx
|
| Data processing |
Fluorescence intensity data has obtained automatically by scanner software. Intensity of signal should subtract the background and normalize to the Positive Control signals before proceeding to analysis. If background is higher than signal, One-step Tukey’s BiWeight (TBW) mehod would be applied. For best results, it is recommended that comparing signal intensities representing the MEDIAN background signals minus local background.
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| |
|
| Submission date |
Oct 06, 2015 |
| Last update date |
Aug 11, 2017 |
| Contact name |
Chien-Wei Lee |
| E-mail(s) |
icehikki@gmail.com
|
| Organization name |
National Yang-Ming University
|
| Street address |
No.155, Sec.2, Linong Street
|
| City |
Taipei |
| ZIP/Postal code |
112 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL20989 |
| Series (1) |
| GSE73762 |
Modulation of reversibility by DNA Methyltransferases during hepatogenic differentiation in Mesenchymal Stromal Cells [protein expression] |
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