|
| Status |
Public on Apr 14, 2022 |
| Title |
HepaRG cells treated with SV-VD3 and OA/PA 2 |
| Sample type |
RNA |
| |
|
| Source name |
Treated with William E complete medium with a mix of oleic acid [200 µM] and palmitic acid [200 µM]+synthetic vitamin D3 [10 nM]
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: hepatic cell line
features: Immortalized hepatic cell line terminally differentiated
age: Adult
|
| Treatment protocol |
At 70% confluence, HepaRG cells were incubated for 24 hrs with either the vehicle or the three different sources of Vitamin D at final concentration of α-Tocopherol in the colture medium of 10 nM. Subsequently HepaRG were incubated with OA/PA mix at the final concentration in the colture medium of 200 µM. Cell treatments with this OA/PA lasted for 48 hrs.
|
| Growth protocol |
Cells were seeded at a density of 10,000 cells/well and kept at 37˚C in a humidified 5% CO2 cell culture incubator.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cells lysates using an QIAsymphony RNA Kit (Qiagen,) with the QIAsymphony SP instrument (Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the standard ThermoFisher Scientific protocol from 6 ug total RNA (GeneChip™ 3’ IVT Pico Kit Manual Workflow).
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| |
|
| Hybridization protocol |
Fragmented and labeled cDNA was prepared according to the manufacturer's instructions and hybridized to a Human Clariom Go Screen 384-array plate for 17 h at 45°C on a GeneTitan MC Instrument (Applied Biosystems, ThermoFisher Scientific). Arrays were washed and stained on a GeneTitan MC Instrument (Applied Biosystems, ThermoFisher Scientific).
|
| Scan protocol |
Arrays were scanned using the GeneTitan MC Instrument (Applied Biosystems, ThermoFisher Scientific). Affymetrix GeneChip Command Console Software (AGCC, ThermoFisher Scientific) was used for GeneTitan MC Instrument control.
|
| Data processing |
Data analysis were performed using Transcriptomic Analysis Console Software (TAC, ThermoFisher Scientific) that provides quality control analysis, performs normalization and summarization, based on the Signal Space Transformation-Robust Multi-Chip Analysis (SST-RMA) analysis algorithm, and provides a list of differentially expressed genes (Limma Bioconductor package, pvalue<0,05) from the biological duplicates for WGO-VE_OA or triplicates of the other condition.
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| |
|
| Submission date |
Apr 13, 2022 |
| Last update date |
Apr 14, 2022 |
| Contact name |
Jacopo Lucci |
| E-mail(s) |
jlucci@biostherapy.it
|
| Phone |
+393428018168
|
| Organization name |
Bios-Therapy, Physiological Systems For Health S.p.A
|
| Lab |
Laboratory of system biology
|
| Street address |
Loc. Aboca, 20
|
| City |
Sansepolcro |
| State/province |
Arezzo |
| ZIP/Postal code |
52037 |
| Country |
Italy |
| |
|
| Platform ID |
GPL31262 |
| Series (1) |
| GSE200765 |
Vitamin D cytoprotection function in human liver cell lipotoxicity and its transcriptomic fingerprint: a comparison of natural and synthetic formulations. |
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