|
| Status |
Public on Mar 24, 2016 |
| Title |
PHH from Donor 1 (D6), treated with DMSO |
| Sample type |
RNA |
| |
|
| Source name |
primary human hepatocytes
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
age: 51
|
| Treatment protocol |
Hepatocytes were isolated and seeded at a density of 4x105 viable cells/well onto BioCoat Collagen I Cellware 12-well culture plates in William’s E Medium, supplemented with 10% FBS, 100 U/ml Pen/Strep, 2 mM L-glutamine,32 mU/ml human insulin, 1 mM sodium pyruvate, 1 x nonessential amino acids, 15 mM HEPES, and 0.8 µg/ml hydrocortisone. After 24 h fully attached cells were adapted for another 12 h to William’s E Medium, supplemented with 10% FBS, 100 U/ml Pen/Strep, 2 mM L-glutamine,32 mU/ml human insulin, 0.1 µM dexamethasone and 0.1% DMSO. Cells were treated for 24 hours in fresh medium with 1µM CITCO, 10 µM rifampicin, 50 µM WY14,643 or vehicle (0.1% dimethylsulfoxide (DMSO).
|
| Growth protocol |
After the treatment for 24h with the indicated agonists the cells were lysed.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Rneasy kit from Qiagen according to the manufacture´s conditions. All (24) used RNA samples had an RNA integrity number (RIN) > 9 (determined with the Bioanalyzer 2100 from Agilent Technologies) to ensure high quality of RNA.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on Human Gene 1.0ST Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip® scanner 7G (Affymetrix).
|
| Description |
Gene expression data from PHH treated with DMSO
|
| Data processing |
After visual inspection of the obtained GeneChip images, the Affymetrix Expression Console (Affymetrix) was used for quality control of microarrays and pre-processing of expression data via robust multi-array average (RMA; Gene Level - Default).
|
| |
|
| Submission date |
Dec 18, 2015 |
| Last update date |
Mar 29, 2016 |
| Contact name |
Ulrich M. Zanger |
| E-mail(s) |
[email protected]
|
| Organization name |
Dr. Margarete Fischer-Bosch-Institute of Clinical Pharmacology
|
| Street address |
Auerbachstr. 112
|
| City |
Stuttgart |
| ZIP/Postal code |
70376 |
| Country |
Germany |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE76148 |
Genome wide comparison of the inducible transcriptomes of CAR, PXR and PPARα in primary human hepatocytes |
|
