|
| Status |
Public on Nov 29, 2011 |
| Title |
DSiPS3factor_5397-33_D12 |
| Sample type |
RNA |
| |
|
| Source name |
DSiPS3factor_5397-33_D12
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: 5397-33
cell type: DS-iPS-T33
days after induction of differentiation: D12
factors using in making ips cells: OCT3/4, SOX2, and KLF4
|
| Growth protocol |
hiPS cells were maintained and passaged weekly on mitomycin C-treated SNL or irradiated MEF feeder cells. To induce hiPS cells to differentiate into hematopoietic cells, we used a coculture system with murine stromal cell line, AGM-S3 cells. Briefly, 10 undifferentiated hiPS cell colonies (each consisting of approximately 1 x 10^3 cells) were picked up, transferred onto each well of 6-well plates (Sumitomo Bakelite Co), which were coated with 25 Gy-irradiated confluent AGM-S3 cells (approximately 1-2 x 10^5 cells), and cultured in hiPS cell maintenance medium for 3 days. Then culture medium was changed to hematopoietic differentiation medium (IMDM, 10% FBS, 3 mM L-glutamine, 5.5 mg ml-1 human transferrin, 50 mg ml-1 ascorbic acid, 1x10^-4 M 2-ME, 1x10^-4 M nonessential amino acids and 100 ng ml^-1 human vascular endothelial growth factor) and replaced everyday for 12 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with SV total RNA isolation system (Promega) and RNA integrity was assessed by a 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
100 ng aliquots of total RNA were amplified and labeled using a Low Input Quick Amp Labeling Kit (Agilent Technologies) according to the manufacturers’ instructions.
|
| |
|
| Hybridization protocol |
600 ng of Cy3-labeled RNA was hybridized to Agilent Whole Human Genome Oligo Microarray
|
| Scan protocol |
The signals were detected by DNA microarray scanner.
|
| Data processing |
The scanned images were extracted with Feature Extraction Software 10.7.3.1 (Agilent Technologies) using default parameters. Subsequent data processing was performed using the GeneSpringGX11.5 software package (Agilent Technologies) and IPA 8.5 (Tomy digital biology).
|
| |
|
| Submission date |
Nov 23, 2011 |
| Last update date |
Nov 29, 2011 |
| Contact name |
Yasuhiro Ebihara |
| E-mail(s) |
ebihara@ims.u-tokyo.ac.jp
|
| Phone |
+81-3-5449-5694
|
| Fax |
+81-3-5449-5428
|
| Organization name |
The Institute of Medical Science, The University of Tokyo
|
| Lab |
Division of Stem Cell Processing
|
| Street address |
4-6-1, Shirokanedai
|
| City |
Minato-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
108-8639 |
| Country |
Japan |
| |
|
| Platform ID |
GPL14550 |
| Series (1) |
| GSE33911 |
Augmented hematopoiesis in induced pluripotent stem cells derived from patients with Down syndrome. |
|
