|
| Status |
Public on Jul 31, 2019 |
| Title |
ACTA2eGFP+ 2 |
| Sample type |
RNA |
| |
|
| Source name |
human iPSCs differentiated towards SMCs
|
| Organism |
Homo sapiens |
| Characteristics |
cell population: ACTA2eGFP+
|
| Treatment protocol |
Cells were dissociated and sorted based on viability (calcein blue+) and GFP expression.
|
| Growth protocol |
Cells were treated with 24 hours of BMP4, ActivinA, VEGF, and FGF2, followed by 72 hours of BMP4, FGF2, and VEGF to reach a KDR+ mesodermal progenitor stage. Cells were then treated with PDGF-BB, TGF-B, and FGF2 to specify towards a smooth muscle-like ACTA2eGFP+ cell.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
QIAzol extraction of total RNA according to the manufacturer's instructions
|
| Label |
biotin
|
| Label protocol |
Biotin labeling was performed using the WT Plus reagent kit (Affymetrix, Santa Clara, CA) according to the manufacturer's protocol.
|
| |
|
| Hybridization protocol |
The labeled, fragmented DNA was hybridized to the Affymetrix Human Gene 2.0 ST Array for 18 hours in a GeneChip Hybridization oven 640 at 45oC with rotation (60 rpm). The hybridized samples were washed and stained using an Affymetrix fluidics station 450.
|
| Scan protocol |
After staining, microarrays were immediately scanned using an Affymetrix GeneArray Scanner 3000 7G Plus.
|
| Description |
Gene expression data from day 30 ACTA2eGFP+ candidate SMCs.
|
| Data processing |
Raw Affymetrix CEL files were normalized to produce Entrez Gene-identifier-specific expression values using the implementation of the Robust Multiarray Average (RMA) in the affy Bioconductor package (version 1.36.1), using R version 2.15.1 and the Brainarray hugene20sthsentrezgcdf R package (version 17.0.0).
|
| |
|
| Submission date |
Jun 10, 2019 |
| Last update date |
Jul 31, 2019 |
| Contact name |
Boston University Microarray and Sequencing Resource |
| E-mail(s) |
msrdata@bu.edu
|
| Organization name |
Boston University
|
| Department |
Microarray and Sequencing Resource
|
| Street address |
72 East Concord Street, E631
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02118 |
| Country |
USA |
| |
|
| Platform ID |
GPL17930 |
| Series (1) |
| GSE132454 |
Global gene expression of human iPSC directed differentation towards smooth muscle cells using an Acta2eGFP human iPSC reporter line |
|
