|
| Status |
Public on Aug 04, 2016 |
| Title |
CiRA00006_iPSC derived endothelial cells_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
ADPKD-specific iPSC derived endothelial cells
|
| Organism |
Homo sapiens |
| Characteristics |
subject/sample source id: CiRA00006
subject status: autosomal dominant polycystic kidney disease (ADPKD) patient
gender: Male
cell type: iPSC derived endothelial cells
|
| Growth protocol |
The differentiation of the iPSCs into vascular cells was carried out as described previously (Sone et al., 2007; Tatsumi et al., 2011). Briefly, undifferentiated human iPSCs were harvested and transferred to a collagen I-coated dish after adjusting the colonies to an appropriate size. On the second day of incubation, the culture medium was replaced with Primate ES medium without basic fibroblast growth factor (bFGF), supplemented with N2 supplement (Invitrogen)/B27 supplement (Invitrogen) and 6-bromoindirubin-3'-oxime (BIO, 5 μM, SIGMA). Thereafter, the cells were incubated for another 3 days, at which time the culture medium was replaced with StemPro-34 SFM (Invitrogen) supplemented with recombinant human vascular endothelial growth factor (VEGF, 50 ng/ml, PeproTech). After another 3–5 days of incubation, FLK1 (+) VE-cadherin (+) and FLK1 (+) VE-cadherin (-) cells were sorted individually using a FACSAria flow cytometer (BD). Sorted FLK1 (+) VE-cadherin (+) cells were confirmed to remain VE-cadherin positive during the subsequent cell culture and analyses. On the other hand, the sorted FLK1 (+) VE-cadherin (-) cells were differentiated into vascular smooth muscle cells after an additional 18-22 days of differentiation on collagen I-coated dishes supplemented with recombinant human platelet-derived growth factor-BB (PDGF-BB, 20 ng/ml, PeproTech).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the miRNeasy Purification kit (Qiagen) following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
biotin
|
| Label protocol |
Labeling, hybridization and scanning were performed as per Affymetrix GeneChip® Whole Transcript (WT) Sense Target Labeling Assay Manual version 4.
|
| |
|
| Hybridization protocol |
Labeling, hybridization and scanning were performed as per Affymetrix GeneChip® Whole Transcript (WT) Sense Target Labeling Assay Manual version 4.
|
| Scan protocol |
Labeling, hybridization and scanning were performed as per Affymetrix GeneChip® Whole Transcript (WT) Sense Target Labeling Assay Manual version 4.
|
| Description |
CiRA00006 EC
|
| Data processing |
Data from RNA expression array experiments in the form of .cel files was normalized, background corrected and summarized using the RMA algorithm implemented in the Affymetrix Expression Console(TM) version 1.0 software.
|
| |
|
| Submission date |
Oct 28, 2015 |
| Last update date |
Aug 04, 2016 |
| Contact name |
Akira Watanabe |
| E-mail(s) |
[email protected]
|
| Organization name |
CyberomiX Inc
|
| Street address |
Rm504, Miyako Bldg, 233, Isa-cho, kamigyo-ku
|
| City |
Kyoto |
| State/province |
Kyoto |
| ZIP/Postal code |
602-8407 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6244 |
| Series (2) |
| GSE74452 |
Identification of a novel risk factor for intracranial aneurysms in ADPKD using iPSC models [Affymetrix] |
| GSE74453 |
Identification of a novel risk factor for intracranial aneurysms in ADPKD using iPSC models |
|
