|
| Status |
Public on Apr 17, 2011 |
| Title |
Fibroblast-iPS, Rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
Foreskin fibroblast-derived iPS cell
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: foreskin fibroblast-derived iPS cell
passage: 5
|
| Treatment protocol |
iPS cells were generated by overexpressing the human transcription factors OCT4, SOX2, C-MYC, KLF4, and NANOG using a doxycycline-inducible lentivirus transgene system that has been previously documented to faithfully reprogram both human fibroblasts and keratinocytes to an hESC-like state (Maherali et al, 2008; Hockemeyer et al., 2008).
|
| Growth protocol |
The hepatocytes were obtained from Lonza (reference #: CC-2703W6). The newborn foreskin (BJ) fibroblasts were obtained from ATCC (reference #: CRL-2522). The melanocytes were obtained from Promocell (reference #: C-12402).
Somatic cells were grown in 425 mls High-glucose DMEM, 50 mls Atlanta FBS (10%), 5 mls L-glutamine, 10 mls Penicillin/Streptomycin, 5 mls Non-essential amino acids, 5 mls Sodium pyruvate, and 2 μl Beta-mercaptoethanol. ES and iPS cells were grown in a hES-KSR Basal Medium of 382.5 mls DMEM/F12 containing Hepes, 100 mls Knockout Serum ment (KSR), 2.5 mls L-glutamine, 5 mls Non-essential amino acids, 10 mls Penicillin/Streptomycin, and 3.5 μl Beta-mercaptoethanol.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy Mini RNA isolation kit (Qiagen).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared using the Affymetrix GeneChip Whole Transcript Sense Target Labeling Assay.
|
| |
|
| Hybridization protocol |
Hybridization was performed according to the Affymetrix GeneChip Whole Transcript Sense Target Labeling Assay Manual.
|
| Scan protocol |
Arrays were scanned with GCS3000 using the standard procedures recommended by Affymetrix.
|
| Description |
Expression profile of human foreskin fibroblast-derived iPS cell (passage number 5).
|
| Data processing |
The data were normalized using RMA in R 2.10.1 and a CDF file that maps the Affymetrix probes to RefSeq genes.
|
| |
|
| Submission date |
Jul 20, 2010 |
| Last update date |
Apr 17, 2011 |
| Contact name |
Jun S Song |
| Organization name |
UCSF
|
| Department |
Institute for Human Genetics
|
| Street address |
513 Parnassus Avenue, Box 0794
|
| City |
San Francisco |
| State/province |
CA |
| ZIP/Postal code |
94143-0794 |
| Country |
USA |
| |
|
| Platform ID |
GPL10666 |
| Series (1) |
| GSE23034 |
Incomplete DNA methylation underlies a transcriptional memory of somatic cells in human iPS cells |
|
