|
| Status |
Public on Sep 15, 2019 |
| Title |
2D HTR8 B |
| Sample type |
RNA |
| |
|
| Source name |
Immortalized human extravillous trophoblast cells
|
| Organism |
Homo sapiens |
| Characteristics |
culture condition: 2D monolayers
tissue: Placenta
|
| Treatment protocol |
96-well ultra-low attachment (ULA) plates (Corning) were used to promote the self-assembly of HTR8/SVneo cells into three-dimensional (3D) cellular spheroids. 2D monolayer samples were simply cultured on flat-bottom plates.
|
| Growth protocol |
HTR8/SVneo cells were cultured at 37°C in a humidified atmosphere of 5% CO2/95% atmospheric air in RPMI-1640 (Corning), supplemented with 5% heat-inactivated FBS (Gibco), 1% (v/v) penicillin-streptomycin (Corning) and 1% (v/v) L-glutamine (Corning). Media was replaced every two days, and at 70-80% confluency, HTR8/SVneo cells were sub-divided.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cells using TRIzol Reagent (Invitrogen) and Direct-zol RNA MiniPrep Kit (Zymo Research), following the manufacturer’s protocol.
|
| Label |
Biotin
|
| Label protocol |
200 ng of RNA was processed and labelled with Biotin Allonamide Triphosphate.
|
| |
|
| Hybridization protocol |
Samples were hybridized for 16 hr at 45°C using Affymetrix WT PLUS Kit.
|
| Scan protocol |
Affymetrix GeneChip™ 3000 instrument system
|
| Data processing |
Normalization and output file generation was performed with Affymetrix Gene Expression Console (Thermo) using the Robust Multi-Chip Analysis (RMA) algorithm [RMA-Gene-full].
|
| |
|
| Submission date |
Feb 21, 2019 |
| Last update date |
Sep 15, 2019 |
| Contact name |
Michael Wong |
| E-mail(s) |
wongm20@mcmaster.ca
|
| Organization name |
McMaster University
|
| Street address |
1200 Main Street West, Health Sciences Building
|
| City |
Hamilton |
| State/province |
Ontario |
| ZIP/Postal code |
L8N 3Z5 |
| Country |
Canada |
| |
|
| Platform ID |
GPL23159 |
| Series (1) |
| GSE126844 |
Transcriptomic analysis of human extravillous trophoblast cells (HTR8/Svneo) cultured as 3D spheroids and 2D monolayers |
|
