|
| Status |
Public on Dec 14, 2012 |
| Title |
Sox17TPd08 |
| Sample type |
RNA |
| |
|
| Source name |
CD34+CD43+CD45low cells/TPEBd08 SOX17-ERT
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CD34+CD43+CD45low hemogenic endothelium-like cells
days of eb differentiation: 8
transduction: transduced with a Sox17-ERT retrovirus cocultured with OP9 stromal cells in the presence of 4OH-tamoxifen
|
| Growth protocol |
ES cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM)-F12 (Sigma) supplemented with 1x MEM non-essential amino acids (Gibco-Invitrogen), 1x GlutaMAX-I (Gibco-Invitrogen), 20% knockout serum replacement (KSR) (Gibco-Invitrogen), 0.1 mM 2-mercaptoethanol (2-ME)(Sigma), and 5 ng/mL basic fibroblast growth factor (bFGF)(ReproCELL) either on irradiated MEF as feeder layers.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
45 μL of fragmented single-stranded DNA was mixed with 15 μL of master mix containing 5X TdT buffer, TdT and DNA Labeling reagent, and was incubated at 37°C for 60 minutes.
|
| |
|
| Hybridization protocol |
The hybridization cocktail was denatured at 99°C for 5 minutes, incubated at 45°C for 5 minutes, and then injected into a GeneChip ST cartridge. The arrays were incubated at 45°C for 17 hours ± 1 hour in a rotating oven at 60 rpm. The arrays were then washed and stained according to the GeneChip Expression Wash, Stain and Scan User Manual (P/N702731).
|
| Scan protocol |
Fluorescence signals were scanned with the Affymetrix GeneChip Scaner 3000. Affymetrix GeneChip Command Console (AGCC) software was used for acquisition of the initial image of the scanned array.
|
| Description |
CD34+CD43+CD45low hemogenic endothelium-like cells transduced with a Sox17-ERT retrovirus cocultured with OP9 stromal cells in the presence of 4OH-tamoxifen
|
| Data processing |
The Affymetrix Expression Console was used to normalize data and determine signal intensity (RMA-Sketch).
|
| |
|
| Submission date |
Apr 17, 2012 |
| Last update date |
Dec 14, 2012 |
| Contact name |
Takaho A. Endo |
| E-mail(s) |
[email protected]
|
| Organization name |
RIKEN
|
| Department |
IMS
|
| Lab |
Laboratory for Integrative Genomics
|
| Street address |
1-7-22 Suehiro, Tsurumi
|
| City |
Yokohama |
| State/province |
Kanagawa |
| ZIP/Postal code |
230-0045 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6244 |
| Series (2) |
| GSE37348 |
Expression data of human ES cells-derived CD34+CD43+CD45low cells (hemogenic endothelium-like cells) expanded upon overexpression of Sox17 |
| GSE38156 |
Critical role of SOX17 in the hematopoietic development from human embryonic stem cells |
|
